) are , , L·mol−1 at 298 K, 303 K, and 308 K, respectively. The enthalpy change () and entropy change () for this reaction are 26.42 kJ·mol−1 and 171.7 J·K−1, respectively. It means that the hydrophobic interaction is the main intermolecular force of the interaction between CHL and Ngb. Synchronous fluorescence spectra showed that the microenvironment of tryptophan and tyrosine residues of Ngb has been changed slightly. The fluorescence quenching efficiency of CHL to tyrosine residues is a little bit more than that to tryptophan residues of Ngb. Furthermore, CD spectra indicated that CHL can induce the formation of α-helix of Ngb.">
Multispectroscopic氯霉素与人类Neuroglobin之间的相互作用的研究</gydF4y2Batitle>
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<h1 class="articleHeader__title">Multispectroscopic氯霉素与人类Neuroglobin之间的相互作用的研究</gydF4y2Bah1>
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<span class="articleHeader__authors_author">Lei黄<gydF4y2Ba!-- -->,<gydF4y2Basup>1</gydF4y2Basup></span>
<span class="articleHeader__authors_author"><strong>Lianzhi李</gydF4y2Bastrong><a href="//www.newsama.com/cdn-cgi/l/email-protection" aria-label="Mail Option"><span role="img" class="anticon">
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</svg></span></a>,<gydF4y2Basup>1</gydF4y2Basup></span>
<span class="articleHeader__authors_author">海丽李<gydF4y2Ba!-- -->,<gydF4y2Basup>1</gydF4y2Basup></span>
<span class="articleHeader__authors_author">Chaohui高<gydF4y2Ba!-- -->,<gydF4y2Basup>1</gydF4y2Basup></span>
<span class="articleHeader__authors_author">回族崔<gydF4y2Ba!-- -->,<gydF4y2Basup>1</gydF4y2Basup></span>
<span class="articleHeader__authors_author">和<gydF4y2Ba!-- -->Xiangshi谭<gydF4y2Basup>1</gydF4y2Basup></span>
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<strong>发表</gydF4y2Bastrong>
<span>2012年6月3日</gydF4y2Baspan>
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<h4 class="header" id="abstract">文摘</gydF4y2Bah4>
<p>氯霉素(背影)和neuroglobin之间的交互(Ngb)调查通过使用荧光,同步荧光、紫外可见和圆二色性(CD)光谱。人们已经发现,背影分子的固有荧光熄灭Ngb在动态猝灭机制的一种方式,通过紫外可见光谱数据的支持。他们的有效猝灭常数(<gydF4y2Basvg height="14.3875" id="M1" style="vertical-align:-3.25792pt;width:26.424999px;" version="1.1" viewbox="0 0 26.424999 14.3875" width="26.424999" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.3875)">
<g transform="translate(72,-60.49)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-62.42,60.66)">
<tspan style="font-size: 8.75px; " x="0" y="0">
年代</gydF4y2Batspan>
<tspan style="font-size: 8.75px; " x="4.691072" y="0">
V</gydF4y2Batspan>
</text>
</g>
</g>
</svg>)<gydF4y2Basvg height="14.375" id="M2" style="vertical-align:-0.3135pt;width:58.325001px;" version="1.1" viewbox="0 0 58.325001 14.375" width="58.325001" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.375)">
<g transform="translate(72,-60.5)">
<text transform="matrix(1,0,0,-1,-71.95,60.86)">
<tspan style="font-size: 12.50px; " x="0" y="0">
2</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="6.2515001" y="0">
。</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.3772497" y="0">
2</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="18.404415" y="0">
×</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="29.182001" y="0">
1</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="35.433502" y="0">
0</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-30.26,66.03)">
<tspan style="font-size: 8.75px; " x="0" y="0">
4</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,<gydF4y2Basvg height="14.375" id="M3" style="vertical-align:-0.3135pt;width:58.325001px;" version="1.1" viewbox="0 0 58.325001 14.375" width="58.325001" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.375)">
<g transform="translate(72,-60.5)">
<text transform="matrix(1,0,0,-1,-71.95,60.86)">
<tspan style="font-size: 12.50px; " x="0" y="0">
2</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="6.2515001" y="0">
。</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.3772497" y="0">
6</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="18.404415" y="0">
×</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="29.182001" y="0">
1</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="35.433502" y="0">
0</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-30.26,66.03)">
<tspan style="font-size: 8.75px; " x="0" y="0">
4</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,<gydF4y2Basvg height="14.375" id="M4" style="vertical-align:-0.3135pt;width:84.8125px;" version="1.1" viewbox="0 0 84.8125 14.375" width="84.8125" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.375)">
<g transform="translate(72,-60.5)">
<text transform="matrix(1,0,0,-1,-71.95,60.86)">
<tspan style="font-size: 12.50px; " x="0" y="0">
一个</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="5.551332" y="0">
n</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="11.802832" y="0">
d</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="21.16758" y="0">
3</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="27.419079" y="0">
。</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="30.544828" y="0">
1</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="39.584499" y="0">
×</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="50.362083" y="0">
1</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="56.613583" y="0">
0</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-9.08,66.03)">
<tspan style="font-size: 8.75px; " x="0" y="0">
4</gydF4y2Batspan>
</text>
</g>
</g>
</svg>L·摩尔<gydF4y2Basup>−1</gydF4y2Basup>在298 K, 303 K, 308 K,分别。焓的变化(<gydF4y2Basvg height="10.6625" id="M5" style="vertical-align:-0.0pt;width:26.4625px;" version="1.1" viewbox="0 0 26.4625 10.6625" width="26.4625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.6625)">
<g transform="translate(72,-63.47)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>)和熵变(<gydF4y2Basvg height="10.8125" id="M6" style="vertical-align:-0.1254pt;width:22.6625px;" version="1.1" viewbox="0 0 22.6625 10.8125" width="22.6625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.8125)">
<g transform="translate(72,-63.35)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>这个反应)26.42 kJ·摩尔<gydF4y2Basup>−1</gydF4y2Basup>和171.7 J·K<gydF4y2Basup>−1</gydF4y2Basup>,分别。这意味着疏水作用的主要分子间作用力的背影和Ngb之间的交互。同步荧光光谱表明,色氨酸和酪氨酸残基的微环境Ngb已经发生了微妙的变化。的背影,酪氨酸残基的荧光猝灭效率比这一点Ngb的色氨酸残基。此外,CD光谱表明,排名可以诱导形成的<gydF4y2Bai>α</gydF4y2Bai>Ngb的螺旋。</gydF4y2Bap>
<span class="end-abs"></span>
<h4 id="introduction">1。介绍</gydF4y2Bah4>
<p>氯霉素(排名2 2-dichloro-N - ((1 r, 2 r) - 1, 3-dihydroxy-1——(4-nitrophenyl) propan-2-yl)乙酰胺,化学结构如图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig1/" target="_blank">1</一个>)是一个广泛的抑菌抗菌;它表现出抗菌活性对革兰氏阳性和革兰氏阴性细菌,以及其他微生物组(<一个href="#B1">1</一个>,<gydF4y2Ba一个href="#B2">2</一个>]。的背影也被证明是有效对抗流行性脑膜炎球菌性脑膜炎(<一个href="#B3">3</一个>];发挥其抗菌效果通过抑制细菌蛋白质合成通过绑定到50年代核糖体亚基干涉的肽基转移酶(<一个href="#B4">4</一个>]。的背影被广泛用于治疗家畜,水产养殖物种,畜牧业和人类消费。然而,背影展示了人类有害的副作用,如致命的再生障碍性贫血,骨髓抑制,和灰色综合症。再生障碍性贫血的发生率在欧洲是每100000人0.2例,然而,在中国,这一数字达到每100000(2例<一个href="#B2">2</一个>,<gydF4y2Ba一个href="#B5">5</一个>]。氯霉素是发现有一个广泛的分布在人类中,如心脏、肺、肾、肝、脾、胸膜液、精液、腹水,和唾液,不管政府的方式<一个href="#B6">6</一个>,<gydF4y2Ba一个href="#B7">7</一个>]。的背影可以穿透血脑屏障,因此在脑脊液浓度相对较高,可达到60%左右的等离子体(<一个href="#B8">8</一个>),而在脑组织,浓度等于甚至高于等离子体(<一个href="#B9">9</一个>]。</gydF4y2Bap>
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<span class="caption-text">图1</gydF4y2Baspan>
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<td>氯霉素的化学结构。</gydF4y2Batd>
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<p>Neuroglobin (Ngb),球蛋白家族的新成员,是最近发现的Burmester等人在人类和小鼠(unfpa<一个href="#B10">10</一个>,<gydF4y2Ba一个href="#B11">11</一个>]。主要表达在微摩尔的集中在几个地区的中枢神经系统,甚至更在视网膜(<一个href="#B10">10</一个>,<gydF4y2Ba一个href="#B11">11</一个>]。尽管其生理功能仍然是不确定的,许多假说提出了关于它的作用在身体Ngb: (i)它是O<gydF4y2Basub>2</gydF4y2Basub>把蛋白促进O<gydF4y2Basub>2</gydF4y2Basub>扩散的细胞需要有氧线粒体呼吸(<一个href="#B10">10</一个>,<gydF4y2Ba一个href="#B12">12</一个>,<gydF4y2Ba一个href="#B13">13</一个>];(2)在缺乏O<gydF4y2Basub>2</gydF4y2Basub>结束,它含NADH氧化酶和提高生产ATP通过糖酵解<一个href="#B14">14</一个>];(3)它充当一个O<gydF4y2Basub>2</gydF4y2Basub>传感器和激活其他蛋白质,有特定的监管职能<一个href="#B15">15</一个>];(iv)可能参与了一氧化氮代谢;(v)它可以保护神经元免受伤害的缺氧和缺血<一个href="#B13">13</一个>]。</gydF4y2Bap>
<p>值得注意的是Ngb主要表达在中枢神经系统中,与此同时我们注意到这一事实的背影可以穿透血脑屏障及其在脑组织浓度等于甚至超过那些在等离子体。考虑所有这些因素,有必要研究之间的交互的背影和Ngb更好地理解他们的分子间相互作用机理和可能的有害副作用的背影,以及Ngb的生理作用。本文的背影和之间的交互Ngb在不同的温度下研究了荧光,紫外-和CD谱,旨在更好地理解的毒理学效应的背影,在分子水平上探讨其绑定机制。</gydF4y2Bap>
<h4 id="experimental">2。实验</gydF4y2Bah4>
<h5 id="sec2.1">2.1。材料和工具</gydF4y2Bah5>
<p>氯霉素是购自Sangon(上海,中国)。pET3a质粒DNA包含人类Ngb的基因就送给教授Burmester, unfpa德国古腾堡大学的美因茨。<gydF4y2Bai>大肠杆菌。</gydF4y2Bai>BL21 (DE3) plysS细胞被保存在我们的实验室。胰蛋白胨和酵母提取物从Oxoid有限公司购买(英格兰)。Isopropyl-1-thio-D-galactopyranoside (IPTG)和二硫苏糖醇(DTT)从σ(美国)。所有其他试剂的分析或生物化学成绩。</gydF4y2Bap>
<p>AKTA净化器100快速蛋白液相色谱(Amersham生物科学、瑞典),HP8453A二极管阵列分光光度计(美国),LS-55荧光光谱仪(美国珀金埃尔默)和Jasco j - 810分光偏振计(日本)。</gydF4y2Bap>
<h5 id="sec2.2">2.2。制备Neuroglobin</gydF4y2Bah5>
<p>pET3a质粒的转化为人类Ngb基因<gydF4y2Bai>大肠杆菌。</gydF4y2Bai>BL21 (DE3) plysS细胞,Ngb表达,纯化进行了如前所述[<一个href="#B16">16</一个>,<gydF4y2Ba一个href="#B17">17</一个>];最后,一个红色的可溶性电泳纯化蛋白。Ngb的浓度原液决心spectrophotometrically通过使用532 nm 10.7更易吸收系数<gydF4y2Basup>−1</gydF4y2Basup>·L·厘米<gydF4y2Basup>−1</gydF4y2Basup>(<一个href="#B18">18</一个>]。</gydF4y2Bap>
<h5 id="sec2.3">2.3。光谱测量</gydF4y2Bah5>
<p>稀释Ngb的股票在20更易·L<gydF4y2Basup>−1</gydF4y2Basup>Na<gydF4y2Basub>2</gydF4y2Basub>HPO<gydF4y2Basub>4</gydF4y2Basub>不<gydF4y2Basub>2</gydF4y2Basub>阿宝<gydF4y2Basub>4</gydF4y2Basub>(pH值7.0)缓冲溶液立即使用前就做好了准备。股票的解决方案(5×10<gydF4y2Basup>−4</gydF4y2Basup>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>)的排名是在20更易·L<gydF4y2Basup>−1</gydF4y2Basup>Na<gydF4y2Basub>2</gydF4y2Basub>HPO<gydF4y2Basub>4</gydF4y2Basub>不<gydF4y2Basub>2</gydF4y2Basub>阿宝<gydF4y2Basub>4</gydF4y2Basub>(pH值7.0)缓冲溶液。</gydF4y2Bap>
<p>荧光光谱被记录在LS-55荧光光谱仪(美国珀金埃尔默)在不同温度(298 K, 303 K, 308 K),与10/5 nm激发/发射通道的带宽。激发波长为280 nm,发射是在290 - 420海里。适当的空格缓冲区被减去对应正确的背景荧光。</gydF4y2Bap>
<p>同步荧光光谱收集同时扫描激发和发射单色仪的波长250 - 600纳米,常数时间间隔(<gydF4y2Basvg height="10.9375" id="M7" style="vertical-align:-0.20064pt;width:54.875px;" version="1.1" viewbox="0 0 54.875 10.9375" width="54.875" xmlns="http://www.w3.org/2000/svg">
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</svg>、60和80 nm,职责)。扫描300海里/分钟的速度和带宽的10/5 nm激发/发射通道。荧光强度是策划和激发波长。</gydF4y2Bap>
<p>在室温下吸收光谱被记录在8453年惠普二极管阵列谱仪(美国惠普公司(hewlett - packard)。</gydF4y2Bap>
<p>圆二色性光谱测量与Jasco j - 810分光偏振计(Jasco、日本)氮以恒定流量5 L / min。技术参数设置是250 - 190 nm波长范围,扫描速率100海里/分钟,通路长度试管0.1厘米,分辨率0.1 nm,时间常数0.1 s和扫描的数量2。</gydF4y2Bap>
<h4 id="results-and-discussion">3所示。结果与讨论</gydF4y2Bah4>
<h5 id="sec3.1">3.1。荧光光谱</gydF4y2Bah5>
<p>芳香族氨基酸的固有荧光蛋白质长期以来一直作为一种有效的方法来研究蛋白质局部构象。色氨酸残基的荧光敏感蛋白质结构的扰动,而量子收益率低苯丙氨酸和酪氨酸渲染这些探针对此类研究不太有用。三个色氨酸和酪氨酸残基四个位于Ngb的不同领域<一个href="#B19">19</一个>]。为了研究绑定Ngb的背影,Ngb的荧光发射光谱的背影在不同浓度的记录在激发280海里;数据在三个温度298 K, 303 K, 308 K收集,分别。图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig2/" target="_blank">2</一个>显示了Ngb的荧光发射光谱的背影在不同浓度的303 K。从图可以看出<一个href="//www.newsama.com/journals/jspec/2012/192591/fig2/" target="_blank">2</一个>Ngb的荧光强度降低而不改变发射波长和峰值形状的背影浓度的增加,建议的背影可以与Ngb及其内在荧光猝灭。</gydF4y2Bap>
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<span class="caption-text">图2</gydF4y2Baspan>
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<td>荧光发射光谱的Ngb在303 K,在缺乏(a)和存在的背影3<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(b) 6<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(c) 9<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(d), 12<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(e), 15<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(f), 18岁<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(g), 21<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(h), 27岁<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(我),33<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(j)和36<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(k);蛋白质的浓度是5<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>。</gydF4y2Batd>
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<p>荧光猝灭过程可以发生在不同的机制,通常划分为动态猝灭和静态猝灭。这可能是杰出的变量依赖于温度和粘度(<一个href="#B20">20.</一个>]。自从在更高的温度下分子的扩散系数为动态猝灭更大了,预计猝灭常数随着温度的增加而增加。相比之下,一个更高的温度会降低其稳定性的复杂,导致较低的淬火为静态猝灭常数(<一个href="#B2">2</一个>]。分析了荧光猝灭通常使用Stern-Volmer方程[<一个href="#B21">21</一个>]<ggydF4y2BaydF4y2Baspan class="displayed-label" id="EEq1">
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<p>在这个方程,<gydF4y2Basvg height="14.3875" id="M9" style="vertical-align:-3.25793pt;width:15.875px;" version="1.1" viewbox="0 0 15.875 14.3875" width="15.875" xmlns="http://www.w3.org/2000/svg">
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0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>和<gydF4y2Basvg height="10.325" id="M10" style="vertical-align:-0.0pt;width:12.05px;" version="1.1" viewbox="0 0 12.05 10.325" width="12.05" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.325)">
<g transform="translate(72,-63.74)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>稳态荧光强度在淬火的缺失和存在物质,分别。<gydF4y2Basvg height="14.3875" id="M11" style="vertical-align:-3.25792pt;width:26.424999px;" version="1.1" viewbox="0 0 26.424999 14.3875" width="26.424999" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.3875)">
<g transform="translate(72,-60.49)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-62.42,60.66)">
<tspan style="font-size: 8.75px; " x="0" y="0">
年代</gydF4y2Batspan>
<tspan style="font-size: 8.75px; " x="4.691072" y="0">
V</gydF4y2Batspan>
</text>
</g>
</g>
</svg>是Stern-Volmer猝灭常数,<gydF4y2Basvg height="16.475" id="M12" style="vertical-align:-4.74141pt;width:14.6625px;" version="1.1" viewbox="0 0 14.6625 16.475" width="14.6625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,16.475)">
<g transform="translate(72,-58.82)">
<text transform="matrix(1,0,0,-1,-71.95,63.6)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-65.17,60.47)">
<tspan style="font-size: 8.75px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>是双分子的猝灭常数,<gydF4y2Basvg height="12.925" id="M13" style="vertical-align:-1.90608pt;width:12.325px;" version="1.1" viewbox="0 0 12.325 12.925" width="12.325" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,12.925)">
<g transform="translate(72,-61.66)">
<text transform="matrix(1,0,0,-1,-71.95,63.61)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>是饮料的浓度<gydF4y2Basvg height="10.875" id="M14" style="vertical-align:-3.25793pt;width:13.125px;" version="1.1" viewbox="0 0 13.125 10.875" width="13.125" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.875)">
<g transform="translate(72,-63.3)">
<text transform="matrix(1,0,0,-1,-71.95,66.6)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-66.42,63.47)">
<tspan style="font-size: 8.75px; " x="0" y="0">
0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>荧光团的生命周期在冷却器的缺失。生物聚合物的荧光寿命是10<gydF4y2Basup>−8</gydF4y2Basup>年代(<一个href="#B22">22</一个>]。的Stern-Volmer曲线<gydF4y2Basvg height="14.75" id="M15" style="vertical-align:-3.25793pt;width:32.150002px;" version="1.1" viewbox="0 0 32.150002 14.75" width="32.150002" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.75)">
<g transform="translate(72,-60.2)">
<text transform="matrix(1,0,0,-1,-71.95,63.5)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-64.22,60.37)">
<tspan style="font-size: 8.75px; " x="0" y="0">
0</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-59.35,63.5)">
<tspan style="font-size: 12.50px; " x="0" y="0">
/</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="3.4758339" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>与[<gydF4y2Basvg height="12.925" id="M16" style="vertical-align:-1.90608pt;width:12.325px;" version="1.1" viewbox="0 0 12.325 12.925" width="12.325" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,12.925)">
<g transform="translate(72,-61.66)">
<text transform="matrix(1,0,0,-1,-71.95,63.61)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>)在不同的温度下被显示在图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig3/" target="_blank">3</一个>,计算<gydF4y2Basvg height="14.3875" id="M17" style="vertical-align:-3.25792pt;width:26.424999px;" version="1.1" viewbox="0 0 26.424999 14.3875" width="26.424999" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.3875)">
<g transform="translate(72,-60.49)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-62.42,60.66)">
<tspan style="font-size: 8.75px; " x="0" y="0">
年代</gydF4y2Batspan>
<tspan style="font-size: 8.75px; " x="4.691072" y="0">
V</gydF4y2Batspan>
</text>
</g>
</g>
</svg>提出了在表值<一个href="//www.newsama.com/journals/jspec/2012/192591/tab1/" target="_blank">1</一个>。图显示,在调查的浓度,结果匹配与Stern-Volmer方程。它也可以从表所示<一个href="//www.newsama.com/journals/jspec/2012/192591/tab1/" target="_blank">1</一个>猝灭常数增加,温度增加,这意味着可能淬火Ngb的排名是一个动态猝灭机制。</gydF4y2Bap>
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<span class="caption-text">表1</gydF4y2Baspan>
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Stern-Volmer猝灭常数<gydF4y2Basvg height="14.3875" id="M18" style="vertical-align:-3.25792pt;width:26.424999px;" version="1.1" viewbox="0 0 26.424999 14.3875" width="26.424999" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.3875)">
<g transform="translate(72,-60.49)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-62.42,60.66)">
<tspan style="font-size: 8.75px; " x="0" y="0">
年代</gydF4y2Batspan>
<tspan style="font-size: 8.75px; " x="4.691072" y="0">
V</gydF4y2Batspan>
</text>
</g>
</g>
</svg>CHL-Ngb体系和热力学参数。</dgydF4y2Baiv>
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<span class="caption-text">图3</gydF4y2Baspan>
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<td>Stern-Volmer情节的淬火Ngb的背影在不同的温度下。</gydF4y2Batd>
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<h5 id="sec3.2">3.2。类型的背影和Ngb之间的相互作用力</gydF4y2Bah5>
<p>药物和生物分子之间的相互作用力量,包括疏水力、氢键、范德华力和静电相互作用[<一个href="#B23">23</一个>]。可以阐明CHL-Ngb交互的绑定模式相互作用的热力学参数。热力学参数的值等<gydF4y2Basvg height="10.6625" id="M24" style="vertical-align:-0.0pt;width:26.4625px;" version="1.1" viewbox="0 0 26.4625 10.6625" width="26.4625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.6625)">
<g transform="translate(72,-63.47)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,<gydF4y2Basvg height="10.85" id="M25" style="vertical-align:-0.15048pt;width:23.1875px;" version="1.1" viewbox="0 0 23.1875 10.85" width="23.1875" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.85)">
<g transform="translate(72,-63.32)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,<gydF4y2Basvg height="10.8125" id="M26" style="vertical-align:-0.1254pt;width:22.6625px;" version="1.1" viewbox="0 0 22.6625 10.8125" width="22.6625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.8125)">
<g transform="translate(72,-63.35)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>可以从以下范霍夫估计方程(<一个href="#B24">24</一个>]。如果温度变化一点点,焓变化(<gydF4y2Basvg height="10.6625" id="M27" style="vertical-align:-0.0pt;width:26.4625px;" version="1.1" viewbox="0 0 26.4625 10.6625" width="26.4625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.6625)">
<g transform="translate(72,-63.47)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>)可以被视为一个常数:<gydF4y2Baspan class="displayed-label" id="EEq3">
<svg height="57.5" id="M28" preserveaspectratio="xMaxYMin" style="vertical-align:-0pt;width:700.9375px;" version="1.1" viewbox="0 0 700.9375 57.5" width="700.9375" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,57.5)">
<g transform="translate(72,-26)">
<text transform="matrix(1,0,0,-1,154.61,56.27)">
<tspan style="font-size: 12.50px; " x="0" y="0">
l</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="3.4758339" y="0">
n</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="11.802832" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="25.806192" y="0">
=</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="39.040001" y="-7.25">
−</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="47.604553" y="-7.25">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="56.631721" y="-7.25">
</gydF4y2Batspan>
</text>
<g>
<path d="m 193.65,59.4 29.63,0" style="fill:none;stroke:#000000;stroke-width:0.82499999;stroke-linecap:butt;stroke-linejoin:miter;stroke-miterlimit:10;stroke-opacity:1;stroke-dasharray:none"></path>
</g>
<text transform="matrix(1,0,0,-1,199.4,47.52)">
<tspan style="font-size: 12.50px; " x="27.860001" y="-8.75">
+</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.2522202" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="40.400002" y="-16">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="49.427166" y="-16">
</gydF4y2Batspan>
</text>
<g>
<path d="m 239.8,59.4 18.02,0" style="fill:none;stroke:#000000;stroke-width:0.82499999;stroke-linecap:butt;stroke-linejoin:miter;stroke-miterlimit:10;stroke-opacity:1;stroke-dasharray:none"></path>
</g>
<text transform="matrix(1,0,0,-1,244.18,47.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="14.84" y="-8.75">
,</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-85.690002" y="17.719999">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-76.662834" y="17.719999">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-63.77224" y="17.719999">
=</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-51.731853" y="17.719999">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-42.704685" y="17.719999">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-29.013901" y="17.719999">
−</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-17.673679" y="17.719999">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-10.79703" y="17.719999">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="-1.769864" y="17.719999">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="6.3570862" y="17.719999">
。</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="220.57001" y="1.65">
(</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="224.73351" y="1.65">
3</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="230.985" y="1.65">
。</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="234.11075" y="1.65">
2</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="240.36224" y="1.65">
)</gydF4y2Batspan>
</text>
</g>
</g>
</svg></span>在(<一个href="#EEq3">3.2</一个>),<ggydF4y2BaydF4y2Basvg height="10.325" id="M29" style="vertical-align:-0.0pt;width:13.2875px;" version="1.1" viewbox="0 0 13.2875 10.325" width="13.2875" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.325)">
<g transform="translate(72,-63.74)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>类似于有效猝灭常数<gydF4y2Basvg height="14.3875" id="M30" style="vertical-align:-3.25792pt;width:26.424999px;" version="1.1" viewbox="0 0 26.424999 14.3875" width="26.424999" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.3875)">
<g transform="translate(72,-60.49)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-62.42,60.66)">
<tspan style="font-size: 8.75px; " x="0" y="0">
年代</gydF4y2Batspan>
<tspan style="font-size: 8.75px; " x="4.691072" y="0">
V</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,<gydF4y2Basvg height="10.325" id="M31" style="vertical-align:-0.0pt;width:11.6875px;" version="1.1" viewbox="0 0 11.6875 10.325" width="11.6875" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.325)">
<g transform="translate(72,-63.74)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>气体常数。焓的变化(<gydF4y2Basvg height="10.6625" id="M32" style="vertical-align:-0.0pt;width:26.4625px;" version="1.1" viewbox="0 0 26.4625 10.6625" width="26.4625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.6625)">
<g transform="translate(72,-63.47)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>)和熵变(<gydF4y2Basvg height="10.8125" id="M33" style="vertical-align:-0.1254pt;width:22.6625px;" version="1.1" viewbox="0 0 22.6625 10.8125" width="22.6625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.8125)">
<g transform="translate(72,-63.35)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>)可以获得线性范霍夫图(图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig4/" target="_blank">4</一个>)。</gydF4y2Bap>
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<span class="caption-text">图4</gydF4y2Baspan>
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<td>范霍夫情节Ngb的互动和合作。</gydF4y2Batd>
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<p>罗斯和萨勃拉曼尼亚(<一个href="#B24">24</一个>)特征相关的热力学参数的符号和大小各种单独的类型的相互作用可能发生在蛋白质协会过程如下:<gydF4y2Baspan class="list"><span class="list-item"><span class="list-label"></span><span class="list-content">
<svg height="11.0625" id="M34" style="vertical-align:-0.30096pt;width:52.25px;" version="1.1" viewbox="0 0 52.25 11.0625" width="52.25" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,11.0625)">
<g transform="translate(72,-63.15)">
<text transform="matrix(1,0,0,-1,-71.95,63.5)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="23.405617" y="0">
></gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="35.446007" y="0">
0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,<gydF4y2Basvg height="11.0625" id="M35" style="vertical-align:-0.30096pt;width:48.775002px;" version="1.1" viewbox="0 0 48.775002 11.0625" width="48.775002" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,11.0625)">
<g transform="translate(72,-63.15)">
<text transform="matrix(1,0,0,-1,-71.95,63.5)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="20.617447" y="0">
></gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="32.657837" y="0">
0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>疏水的力量,</gydF4y2Baspan></span><span class="list-item"><span class="list-label"></span><span class="list-content">
<svg height="11.0625" id="M36" style="vertical-align:-0.30096pt;width:52.25px;" version="1.1" viewbox="0 0 52.25 11.0625" width="52.25" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,11.0625)">
<g transform="translate(72,-63.15)">
<text transform="matrix(1,0,0,-1,-71.95,63.5)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="23.405617" y="0">
<</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="35.446007" y="0">
0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,<gydF4y2Basvg height="11.0625" id="M37" style="vertical-align:-0.30096pt;width:48.775002px;" version="1.1" viewbox="0 0 48.775002 11.0625" width="48.775002" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,11.0625)">
<g transform="translate(72,-63.15)">
<text transform="matrix(1,0,0,-1,-71.95,63.5)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="20.617447" y="0">
<</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="32.657837" y="0">
0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>氢键或范德华力,</gydF4y2Baspan></span><span class="list-item"><span class="list-label"></span><span class="list-content">
<svg height="12.3" id="M38" style="vertical-align:-1.29163pt;width:52.25px;" version="1.1" viewbox="0 0 52.25 12.3" width="52.25" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,12.3)">
<g transform="translate(72,-62.16)">
<text transform="matrix(1,0,0,-1,-71.95,63.5)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="23.405617" y="0">
≤</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="35.446007" y="0">
0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,<gydF4y2Basvg height="11.0625" id="M39" style="vertical-align:-0.30096pt;width:48.775002px;" version="1.1" viewbox="0 0 48.775002 11.0625" width="48.775002" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,11.0625)">
<g transform="translate(72,-63.15)">
<text transform="matrix(1,0,0,-1,-71.95,63.5)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="20.617447" y="0">
></gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="32.657837" y="0">
0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>静电力。</gydF4y2Baspan></span></span></p>
<p>从表可以看出<一个href="//www.newsama.com/journals/jspec/2012/192591/tab1/" target="_blank">1</一个>自由能变化为负(<gydF4y2Basvg height="10.85" id="M40" style="vertical-align:-0.15048pt;width:23.1875px;" version="1.1" viewbox="0 0 23.1875 10.85" width="23.1875" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.85)">
<g transform="translate(72,-63.32)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>< 0),这意味着的背影和Ngb之间的交互是一个自发的过程。和焓值的变化(<gydF4y2Basvg height="10.6625" id="M41" style="vertical-align:-0.0pt;width:26.4625px;" version="1.1" viewbox="0 0 26.4625 10.6625" width="26.4625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.6625)">
<g transform="translate(72,-63.47)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>)和熵变(<gydF4y2Basvg height="10.8125" id="M42" style="vertical-align:-0.1254pt;width:22.6625px;" version="1.1" viewbox="0 0 22.6625 10.8125" width="22.6625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.8125)">
<g transform="translate(72,-63.35)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>)被发现是26.42 kJ·摩尔<gydF4y2Basup>−1</gydF4y2Basup>和171.7 J·摩尔<gydF4y2Basup>−1</gydF4y2Basup>·K<gydF4y2Basup>−1</gydF4y2Basup>分别表示,这个反应是一个吸热作用,疏水作用是主要的约束力的交互。因为<gydF4y2Basvg height="10.8125" id="M43" style="vertical-align:-0.1254pt;width:22.6625px;" version="1.1" viewbox="0 0 22.6625 10.8125" width="22.6625" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.8125)">
<g transform="translate(72,-63.35)">
<text transform="matrix(1,0,0,-1,-71.95,63.52)">
<tspan style="font-size: 12.50px; " x="0" y="0">
Δ</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.0271664" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>> 0,我们也可以推测,绑定是一个熵驱动的自发过程。</gydF4y2Bap>
<h5 id="sec3.3">3.3。绑定平衡分析</gydF4y2Bah5>
<p>当小分子绑定独立一组等效高分子网站,结合常数<gydF4y2Basvg height="10.325" id="M44" style="vertical-align:-0.0pt;width:13.2875px;" version="1.1" viewbox="0 0 13.2875 10.325" width="13.2875" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.325)">
<g transform="translate(72,-63.74)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>和结合位点的数量<gydF4y2Basvg height="7.1374998" id="M45" style="vertical-align:-0.10033pt;width:7.8874998px;" version="1.1" viewbox="0 0 7.8874998 7.1374998" width="7.8874998" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,7.1375)">
<g transform="translate(72,-66.29)">
<text transform="matrix(1,0,0,-1,-71.95,66.44)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>可以通过使用以下方程计算(<一个href="#B25">25</一个>]:<gydF4y2Baspan class="displayed-label" id="EEq4">
<svg height="32.849998" id="M46" preserveaspectratio="xMaxYMin" style="vertical-align:-0pt;width:700.9375px;" version="1.1" viewbox="0 0 700.9375 32.849998" width="700.9375" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,32.85)">
<g transform="translate(72,-45.72)">
<text transform="matrix(1,0,0,-1,135.95,54.52)">
<tspan style="font-size: 12.50px; " x="10.93" y="-9.2700005">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="0" y="0">
l</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="3.4758339" y="0">
g</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,154.6,60.66)">
<tspan style="font-size: 8.75px; " x="0" y="0">
0</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,162.25,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
−</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="11.340221" y="0">
</gydF4y2Batspan>
</text>
<g>
<path d="m 146.88,57.65 36.26,0" style="fill:none;stroke:#000000;stroke-width:0.82499999;stroke-linecap:butt;stroke-linejoin:miter;stroke-miterlimit:10;stroke-opacity:1;stroke-dasharray:none"></path>
</g>
<text transform="matrix(1,0,0,-1,160.24,45.77)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="93.089996" y="-8.7200003">
(</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="97.25" y="-8.75">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="107.02" y="-8.7200003">
]</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="27.57" y="-8.75">
=</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="39.597885" y="-8.75">
l</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="43.073719" y="-8.75">
g</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="51.413219" y="-8.75">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="63.716171" y="-8.75">
+</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="75.056396" y="-8.75">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="81.270386" y="-8.75">
l</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="84.746216" y="-8.75">
g</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="113.27" y="-8.75">
,</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="304.51001" y="-8.75">
(</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="308.67349" y="-8.75">
3</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="314.92499" y="-8.75">
。</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="318.05075" y="-8.75">
3</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="324.30225" y="-8.75">
)</gydF4y2Batspan>
</text>
</g>
</g>
</svg></span>在哪里<gydF4y2Basvg height="14.3875" id="M47" style="vertical-align:-3.25793pt;width:15.875px;" version="1.1" viewbox="0 0 15.875 14.3875" width="15.875" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.3875)">
<g transform="translate(72,-60.49)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-64.22,60.66)">
<tspan style="font-size: 8.75px; " x="0" y="0">
0</gydF4y2Batspan>
</text>
</g>
</g>
</svg>和<gydF4y2Basvg height="10.325" id="M48" style="vertical-align:-0.0pt;width:12.05px;" version="1.1" viewbox="0 0 12.05 10.325" width="12.05" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.325)">
<g transform="translate(72,-63.74)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>是荧光强度之前和之后的饮料,然后呢<gydF4y2Basvg height="12.9875" id="M49" style="vertical-align:-1.95624pt;width:22.737499px;" version="1.1" viewbox="0 0 22.737499 12.9875" width="22.737499" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,12.9875)">
<g transform="translate(72,-61.61)">
<text transform="matrix(1,0,0,-1,-71.95,63.61)">
<tspan style="font-size: 12.50px; " x="0" y="0">
(</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="4.1634989" y="0">
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<tspan style="font-size: 12.50px; " x="13.928342" y="0">
]</gydF4y2Batspan>
</text>
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</g>
</svg>是总弄熄的浓度。图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig5/" target="_blank">5</一个>的情节是<gydF4y2Basvg height="14.8625" id="M50" style="vertical-align:-3.25793pt;width:82.025002px;" version="1.1" viewbox="0 0 82.025002 14.8625" width="82.025002" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.8625)">
<g transform="translate(72,-60.11)">
<text transform="matrix(1,0,0,-1,-71.95,63.41)">
<tspan style="font-size: 12.50px; " x="0" y="0">
l</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="3.4758339" y="0">
g</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.727334" y="0">
(</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="13.890833" y="0">
</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-50.33,60.28)">
<tspan style="font-size: 8.75px; " x="0" y="0">
0</gydF4y2Batspan>
</text>
<text transform="matrix(1,0,0,-1,-42.68,63.41)">
<tspan style="font-size: 12.50px; " x="0" y="0">
−</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="11.340221" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="19.067076" y="0">
)</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="23.230574" y="0">
/</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="26.706408" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>与<gydF4y2Basvg height="14.2" id="M51" style="vertical-align:-2.7337pt;width:34.900002px;" version="1.1" viewbox="0 0 34.900002 14.2" width="34.900002" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,14.2)">
<g transform="translate(72,-60.64)">
<text transform="matrix(1,0,0,-1,-71.95,63.41)">
<tspan style="font-size: 12.50px; " x="0" y="0">
l</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="3.4758339" y="0">
g</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="9.727334" y="0">
(</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="13.890833" y="0">
</gydF4y2Batspan>
<tspan style="font-size: 12.50px; " x="23.655676" y="0">
]</gydF4y2Batspan>
</text>
</g>
</g>
</svg>,并结合位点的数量<gydF4y2Basvg height="7.1374998" id="M52" style="vertical-align:-0.10033pt;width:7.8874998px;" version="1.1" viewbox="0 0 7.8874998 7.1374998" width="7.8874998" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,7.1375)">
<g transform="translate(72,-66.29)">
<text transform="matrix(1,0,0,-1,-71.95,66.44)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>并结合常数<gydF4y2Basvg height="10.325" id="M53" style="vertical-align:-0.0pt;width:13.2875px;" version="1.1" viewbox="0 0 13.2875 10.325" width="13.2875" xmlns="http://www.w3.org/2000/svg">
<g transform="matrix(1.25,0,0,-1.25,0,10.325)">
<g transform="translate(72,-63.74)">
<text transform="matrix(1,0,0,-1,-71.95,63.79)">
<tspan style="font-size: 12.50px; " x="0" y="0">
</gydF4y2Batspan>
</text>
</g>
</g>
</svg>在298 K被发现是1.08和6.3×10<gydF4y2Basup>4</gydF4y2Basup>L·摩尔<gydF4y2Basup>−1</gydF4y2Basup>分别,这表明只有一个类的结合位点在Ngb的背影,和相关系数<gydF4y2Basvg height="10.325" id="M54" style="vertical-align:-0.0pt;width:11.6875px;" version="1.1" viewbox="0 0 11.6875 10.325" width="11.6875" xmlns="http://www.w3.org/2000/svg">
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</svg>(0.997)的背影和Ngb比赛意味着之间的交互与site-binding模型中描述(<一个href="#EEq4">3.3</一个>)。</gydF4y2Bap>
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<span class="caption-text">图5</gydF4y2Baspan>
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<td>双对数图的排名在298 K Ngb荧光猝灭效应。</gydF4y2Batd>
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<h5 id="sec3.4">3.4。同步荧光光谱</gydF4y2Bah5>
<p>同步荧光光谱是一个非常有用的方法来研究氨基酸残基的微环境(<一个href="#B26">26</一个>]。通过选择不同的波长间隔,色氨酸和酪氨酸残基的特征荧光峰重叠的常规荧光光谱可以孤立<一个href="#B27">27</一个>]。如图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig6/" target="_blank">6</一个>,当<gydF4y2Basvg height="10.6625" id="M55" style="vertical-align:-0.0pt;width:11.4125px;" version="1.1" viewbox="0 0 11.4125 10.6625" width="11.4125" xmlns="http://www.w3.org/2000/svg">
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Δ</gydF4y2Batspan>
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</svg><i>λ</gydF4y2Bai>是20 nm, Ngb的荧光发射峰在300海里,这是酪氨酸残基的发射峰<一个href="#B28">28</一个>]。当<gydF4y2Basvg height="10.6625" id="M56" style="vertical-align:-0.0pt;width:11.4125px;" version="1.1" viewbox="0 0 11.4125 10.6625" width="11.4125" xmlns="http://www.w3.org/2000/svg">
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Δ</gydF4y2Batspan>
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</svg><i>λ</gydF4y2Bai>60 nm,小峰在295 nm,大峰在340纳米荧光光谱观察,使酪氨酸和色氨酸残基的特征信息。此外,当<gydF4y2Basvg height="10.6625" id="M57" style="vertical-align:-0.0pt;width:11.4125px;" version="1.1" viewbox="0 0 11.4125 10.6625" width="11.4125" xmlns="http://www.w3.org/2000/svg">
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Δ</gydF4y2Batspan>
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</svg><i>λ</gydF4y2Bai>增加到80纳米,这两个峰的强度增加,和峰值波长转移到311 nm和355 nm,分别;后者被认为是色氨酸残基的发射峰<一个href="#B28">28</一个>]。此外,没有明显的峰值发射波长和形状变化在这些同步荧光光谱,它支持动态猝灭方式之间的交互Ngb的背影。此外,我们比较了荧光猝灭效率排名个人酪氨酸和色氨酸残基的Ngb(图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig7/" target="_blank">7</一个>);结果表明,酪氨酸的荧光猝灭效率排名是比这更明显的色氨酸。</gydF4y2Bap>
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<span class="caption-text">图6</gydF4y2Baspan>
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<td>同步荧光光谱Ngb (pH = 7.0, T = 298 k)没有(a)和存在的背影5<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(b) 8<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(c), 10<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(d), 20<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(e), 30岁<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(f)和40<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(g);蛋白质的浓度是5<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>。(一)<gydF4y2Basvg height="10.6625" id="M58" style="vertical-align:-0.0pt" version="1.1" viewbox="0 0 11.4125 10.6625" width="11.4125" xmlns="http://www.w3.org/2000/svg">
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</svg><i><i>λ</gydF4y2Bai></i>= 20 nm, (B)<gydF4y2Basvg height="10.6625" id="M59" style="vertical-align:-0.0pt" version="1.1" viewbox="0 0 11.4125 10.6625" width="11.4125" xmlns="http://www.w3.org/2000/svg">
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</svg><i><i>λ</gydF4y2Bai></i>= 60 nm, (C)<gydF4y2Basvg height="10.6625" id="M60" style="vertical-align:-0.0pt" version="1.1" viewbox="0 0 11.4125 10.6625" width="11.4125" xmlns="http://www.w3.org/2000/svg">
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Δ</gydF4y2Batspan>
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</svg><i><i>λ</gydF4y2Bai></i>= 80海里。</gydF4y2Batd>
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<span class="caption-text">图7</gydF4y2Baspan>
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<td>荧光猝灭效率的背影Ngb的色氨酸和酪氨酸残基。</gydF4y2Batd>
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<h5 id="sec3.5">3.5。紫外可见光谱</gydF4y2Bah5>
<p>紫外可见光谱研究是一个非常简单的方法来探索一种蛋白质样品的结构变化。图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig8/" target="_blank">8</一个>显示的背影Ngb的紫外可见光谱的影响。Ngb的铁形式显示大的振幅<gydF4y2Bai>α</gydF4y2Bai>乐队(533海里),俗乐队(413海里)<一个href="#B16">16</一个>]。乐队可以分配给为中心280海里<gydF4y2Bai>π</gydF4y2Bai>- - - - - -<gydF4y2Bai>π</gydF4y2Bai>*电子跃迁的色氨酸和酪氨酸残基。在相同的实验条件下,没有观察到吸光度的背影。结果表明,吸光度在413海里只略有下降,但没有峰转变,这表明的背影只有一点点影响Ngb血红素中心分子的微环境和维护其hexacoordinated配置。此外,没有观察到明显的变化在280 nm的吸光度的增加数量的背影。紫外可见光谱结果支持动态猝灭机制通过荧光光谱。</gydF4y2Bap>
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<span class="caption-text">图8</gydF4y2Baspan>
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<td>紫外可见光谱Ngb在缺乏(a)和排名36的存在<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(b), 60<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(c), 84年<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(d), 120年<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(e), 180年<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(f)。使用的蛋白浓度是5<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>pH值7.0,298 K。</gydF4y2Batd>
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<h5 id="sec3.6">3.6。圆二色性谱</gydF4y2Bah5>
<p>圆二色性光谱是一种独特的光学技术研究蛋白质的二级结构的变化。为了研究Ngb的背影对二级结构的影响,CD谱了。图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig9/" target="_blank">9</一个>显示的CD光谱Ngb在缺乏,在不同浓度的背影。Ngb的CD光谱表现出两个负面乐队在208和222海里远紫外区域,导致了<gydF4y2Basvg height="11.7375" id="M61" style="vertical-align:-0.16302pt;width:52.099998px;" version="1.1" viewbox="0 0 52.099998 11.7375" width="52.099998" xmlns="http://www.w3.org/2000/svg">
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</gydF4y2Batspan>
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→</gydF4y2Batspan>
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∗</gydF4y2Batspan>
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</svg>转让的债券的特点<gydF4y2Bai>α</gydF4y2Bai>螺旋结构的蛋白质(<一个href="#B25">25</一个>,<gydF4y2Ba一个href="#B29">29日</一个>]。如图<一个href="//www.newsama.com/journals/jspec/2012/192591/fig9/" target="_blank">9</一个>、峰值强度在222 nm的背影,随着浓度的增加,但没有造成重大转变的峰值。计算结果显示增加了<gydF4y2Bai>α</gydF4y2Bai>螺旋含量从52.6%到64.9%的摩尔比率Ngb的背影1:8日表示,Ngb之间的互动和合作会影响分子内氢绑定交互Ngb的疏水性,与荧光的结果一致。</gydF4y2Bap>
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<span class="caption-text">图9</gydF4y2Baspan>
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<td>CD光谱Ngb在298 K,在缺乏(a)和存在的背影10<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(b), 30岁<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(c)和40<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>(d);蛋白质的浓度是5<gydF4y2Bai>μ</gydF4y2Bai>摩尔·L<gydF4y2Basup>−1</gydF4y2Basup>。</gydF4y2Batd>
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<h4 id="conclusions">4所示。结论</gydF4y2Bah4>
<p>的背影与Ngb的相互作用已经被不同的光谱方法研究包括荧光光谱、同步荧光光谱、紫外可见吸收光谱,CD光谱。荧光光谱结果表明,排名可以通过动态猝灭Ngb的固有荧光猝灭机制,由紫外可见光谱的结果,还支持和相互作用的热力学参数表明,疏水的力量在这个过程中扮演着重要的角色。此外,同步荧光光谱表明,色氨酸和酪氨酸残基的微环境改变,和酪氨酸残基的荧光猝灭效率排名更明显比Ngb的色氨酸残基。此外,CD光谱表明,排名可以诱导形成的<gydF4y2Bai>α</gydF4y2Bai>Ngb的螺旋是一致的背影和Ngb疏水相互作用的结果。</gydF4y2Bap>
<h4 id="acknowledgments">确认</gydF4y2Bah4>
<p>作者感谢教授t Burmester提供人类的基因unfpa neuroglobin友好。这项工作得到了国家自然科学基金(21142003,21142003),中国山东省自然科学基金(没有。ZR2011BL002),山东省泰山学者研究基金。</gydF4y2Bap>
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