. We then created HIP1 mutants to probe our hypothesis that a short hydrophobic path in the opened region is the binding site for clathrin light chain. We found that the binding of hub/LCb was sensitive to mutating two distantly separated basic residues (K474 and K494). The basic patches marked by K474 and K494 are conserved in HIP12/1R. The lack of conservation in sla2p (S. cerevisiae), HIP1 from D. melanogaster, and HIP1 homolog ZK370.3 from C. elegans implies the binding of HIP1 and HIP1 homologs to clathrin light chain may be different in these organisms.">
远离间隔的两个基本的补丁在弹性域Huntingtin-Interacting蛋白1 (HIP1)网格蛋白轻链的结合至关重要</gydF4y2Batitle>
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.expand_btn{-webkit-flex:1;-ms-flex:1;flex:1;background:url('data:image/svg+xml;base64,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') no-repeat 6px;background-position:center;margin-top:10px;min-width:55px;display:none;} @media (min-width:768px){.hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption .expand_btn{display:none;}} .hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-text{-webkit-flex:9;-ms-flex:9;flex:9;text-align:left;padding:21px 37px 10px 0px;font-size:16px;font-weight:700;} @media (min-width:768px){.hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-text{padding:19px 18px 10px 18px;}} @media (min-width:992px){.hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-text{padding:19px 21px 10px 21px;}} .hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-partial-url{-webkit-flex:1;-ms-flex:1;flex:1;background:url('data:image/svg+xml;base64,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') no-repeat 6px;background-position:center;padding:16px 0 16px 0;min-width:62px;max-width:62px;} @media (min-width:768px){.hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-partial-url{padding:16px 0 17px 0;min-width:52px;max-width:52px;}} @media (min-width:992px){.hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-partial-url{padding:15px 0 17px 0;min-width:60px;max-width:60px;}} .hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption .caption-partial-url:hover{-webkit-text-decoration:none;text-decoration:none;} .hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption-text{font-size:14px;padding:0px 18px 17px 22px;} @media (min-width:768px){.hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption-text{padding:0px 15px 15px 18px;}} @media (min-width:992px){.hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption-text{padding:0px 18px 19px 21px;}} .hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption-text figcaption{margin:0;padding:0;} .hMPMVR .floats-partial-section .floats-partial-footer .floats-partial-caption-text svg{max-width:100%;} .hMPMVR ol{font-size:16px;padding-left:32px;font-family:STIXGeneral-Regular;line-height:24px;margin:15px 0;} .hMPMVR ol li{margin:10px 0;padding-left:10px;word-break:normal;} .hMPMVR ol li a{word-break:break-all;} .hMPMVR ol li .reflinks{display:block;} .hMPMVR ol li .reflinks a{color:#4D8A17;-webkit-text-decoration:none;text-decoration:none;cursor:pointer;} .hMPMVR ol li .reflinks a:hover{-webkit-text-decoration:underline;text-decoration:underline;} .hMPMVR ol li .reflinks .sep{color:#000000;} .hMPMVR .table-wrapper{width:auto;overflow-x:auto;} .hMPMVR .table-wrapper::-webkit-scrollbar{-webkit-appearance:none;width:5px;height:5px;} .hMPMVR .table-wrapper::-webkit-scrollbar-thumb{border-radius:5px;background-color:rgba(0,0,0,.5);-webkit-box-shadow:0 0 1px rgba(255,255,255,.5);} @media (min-width:768px){.hMPMVR .table-wrapper{overflow-x:auto;width:auto;-webkit-break-word:word-break;break-word:word-break;margin:0 auto;}} @media (min-width:992px){.hMPMVR .table-wrapper{width:auto;}} @media (min-width:1324px){.hMPMVR .table-wrapper{width:auto;}} @media (min-width:320px){.hMPMVR{margin-bottom:33px;padding:0;}} @media (min-width:992px){.hMPMVR{padding:0;margin-bottom:100px;}} @media (min-width:1325px){.hMPMVR{margin-bottom:100px;}} .hMPMVR .owl-stage{display:-webkit-box;display:-webkit-flex;display:-ms-flexbox;display:flex;} .hMPMVR .floats-partial-middle--thumbs{display:-webkit-box;display:-webkit-flex;display:-ms-flexbox;display:flex;} .hMPMVR .floats-partial-middle--thumbs .partial-carousel-dot svg{fill:#4D8A17;} .hMPMVR .floats-partial-middle--thumbs .partial-carousel-dot strong{color:#4D8A17;font-weight:500;} .hMPMVR .floats-partial-middle--thumbs .partial-carousel-dot.active strong{color:#007B8B;} .hMPMVR 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<p class="sc-kZmsYB bkiEfx article_type">研究信|<gydF4y2Ba!-- -->开放获取</pgydF4y2Ba>
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体积<gydF4y2Ba!-- -->
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<p>乔尔·a·状态,玛丽·e·克莱格,梅丽莎·伊林沃思,克莱尔·冈萨雷斯,钱其琛妞妞<gydF4y2Ba!-- -->,<gydF4y2Ba!-- -->
<!-- -->”<gydF4y2Baspan class="adjust-article-svg-size">远离间隔的两个基本的补丁在弹性域Huntingtin-Interacting蛋白1 (HIP1)网格蛋白轻链的结合至关重要</gydF4y2Baspan>”,<gydF4y2Ba!-- --><i>生物化学研究国际</gydF4y2Bai>,<gydF4y2Ba!-- -->
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<h1 class="sc-iRbamj jZOxOO sc-iSDuPN hHHqqR article_title adjust-article-svg-size heading-title">远离间隔的两个基本的补丁在弹性域Huntingtin-Interacting蛋白1 (HIP1)网格蛋白轻链的结合至关重要</gydF4y2Bah1>
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<span class=""><b>乔尔·a .状态</gydF4y2Bab><a class="sc-htpNat bUhGXt link" href="mailto:" aria-label="Mail Option"><span class="sc-htoDjs kNoiZu sc-hjRWVT irNWJI icon-colored _2zt-1j0LmaF8UjTQAM413G icon-themeable" title="" font-size="16"></span></a>,<gydF4y2Basup>1</gydF4y2Basup></span>
<span class="">玛丽·e·克莱格<gydF4y2Ba!-- -->,<gydF4y2Basup>1</gydF4y2Basup></span>
<span class="">梅丽莎·伊林沃思<gydF4y2Ba!-- -->,<gydF4y2Basup>1</gydF4y2Basup></span>
<span class="">克莱尔·冈萨雷斯<gydF4y2Ba!-- -->,<gydF4y2Basup>1</gydF4y2Basup>
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<span class="">钱妞妞<gydF4y2Basup>1</gydF4y2Basup></span>
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<p><sup>1</gydF4y2Basup><span>印第安纳大学生物学系,Simon大厅405 b, 212 s .霍桑驱动器,布卢明顿,在47405年,美国</gydF4y2Baspan></p>
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<b>学术编辑器:</gydF4y2Bab>William s .特林布尔</gydF4y2Badiv>
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<span class="articleContent__PublicationField-sc-2yl9jy-6 cIribm">接受</gydF4y2Baspan>
<span class="articleContent__PublicationValue-sc-2yl9jy-7 khHUcu">2009年2月24日</gydF4y2Baspan>
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<span class="articleContent__PublicationField-sc-2yl9jy-6 cIribm">发表</gydF4y2Baspan>
<span class="articleContent__PublicationValue-sc-2yl9jy-7 khHUcu">07年4月2009年</gydF4y2Baspan>
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<h4 class="header" id="abstract">文摘</gydF4y2Bah4>
<p>臀部家族蛋白质之间的相互作用(HIP1和HIP12/1R)和网格蛋白是内吞作用的基础。我们用圆二色性(CD)的稳定性研究HIP1 subfragment舒展开放(aa468 - 530)。乳糜泻热融化显示HIP1 468 - 530只在低温下是稳定的,但这HIP1片段包含一个结构单元,甚至在83年不融化<gydF4y2Basvg height="10.9125" id="M1" style="vertical-align:-0.17554pt;width:15px;" version="1.1" viewbox="0 0 15 10.9125" width="15" xmlns="http://www.w3.org/2000/svg">
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</svg>。然后我们创建HIP1突变体探测我们的假设,一个简短的疏水性路径打开地区是网格蛋白轻链的结合位点。我们发现变异中心/ LCb敏感的绑定两个冷淡地分离基本残留物(K474和K494)。基本的补丁,K474 HIP12/1R K494是守恒的。缺乏保护<gydF4y2Bai>sla2p(酵母)</gydF4y2Bai>,HIP1<gydF4y2Bai>d .腹</gydF4y2Bai>,HIP1 ZK370.3同系物<gydF4y2Bai>秀丽隐杆线虫</gydF4y2Bai>意味着HIP1绑定和HIP1同系物,网格蛋白轻链在这些生物可能不同。</pgydF4y2Ba>
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<h4 id="introduction">1。介绍</gydF4y2Bah4>
<p>Huntingtin-interacting蛋白1 (HIP1)及其相对HIP12/1R,出芽的clathrin-coated囊泡(闭路)[<一个href="#B1">1</一个>,<一个href="#B2">2</一个>]。酵母HIP1同族体,<gydF4y2Bai>sla2p</gydF4y2Bai>闭路的发展所需,是酵母(<一个href="#B3">3</一个>,<一个href="#B4">4</一个>]。有共享区域HIP1和HIP12/1R调解网格蛋白的结合,但也有网站每个髋关节特有的蛋白质。例如,HIP1,但不是HIP12/1R,网格蛋白框主题(<gydF4y2Basvg height="14.5375" id="M2" style="vertical-align:-3.25793pt;width:77.0625px;" version="1.1" viewbox="0 0 77.0625 14.5375" width="77.0625" xmlns="http://www.w3.org/2000/svg">
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</svg>)函数绑定氨基β螺旋桨域网格蛋白(<一个href="#B5">5</一个>]。HIP1和HIP12/1R不与适配器蛋白2 (AP2)以同样的方式。的AP2-binding FXDXF和DPF图案(X表示任何氨基酸)只有在HIP1 [<一个href="#B5">5</一个>]。HIP12/1R显然具有较高的亲和力f -肌动蛋白(<一个href="#B6">6</一个>),这表明HIP1 HIP12/1R扮演不同的角色,闭路的形成。许多氨基酸(<gydF4y2Basvg height="14.5375" id="M3" style="vertical-align:-3.25793pt;width:26.7125px;" version="1.1" viewbox="0 0 26.7125 14.5375" width="26.7125" xmlns="http://www.w3.org/2000/svg">
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</svg>KN地区)的卷曲螺旋域HIP1和HIP12/1R已确定影响绑定的网格蛋白轻链(CLC) [<一个href="#B7">7</一个>),但我们不知道<gydF4y2Basvg height="14.5375" id="M6" style="vertical-align:-3.25793pt;width:26.7125px;" version="1.1" viewbox="0 0 26.7125 14.5375" width="26.7125" xmlns="http://www.w3.org/2000/svg">
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<g transform="translate(72,-60.37)">
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<text transform="matrix(1,0,0,-1,-63.61,60.55)">
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</svg>构成整个CLC约束力的决定因素在臀部的蛋白质。我们最近决定两个相邻的晶体结构片段的HIP1卷曲螺旋域(PDB文件:2 no2和2 qa7)跨网格蛋白光chain-binding地区(<一个href="#B8">8</一个>,<一个href="#B9">9</一个>]。DL的<gydF4y2Basvg height="14.5375" id="M8" style="vertical-align:-3.25793pt;width:54.162498px;" version="1.1" viewbox="0 0 54.162498 14.5375" width="54.162498" xmlns="http://www.w3.org/2000/svg">
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</svg>KN拉伸嵌入式舒展开放地区,我们首次发现在HIP1 482 - 586 (2 no2) [<一个href="#B8">8</一个>),后来发现HIP1 371 - 481 (2 qa7) (<一个href="#B9">9</一个>]。在本研究的信我们定义的固有稳定性包含了一段HIP1地区和问静电相互作用推动CLC的绑定。我们生成一个HIP1 subfragment (aa468 - 530)横贯开放地区的圆二色性(CD)实验来评估该域的内在稳定性。然后,我们研究了如果两个冷淡地间隔带正电的补丁打开地区CLC绑定中扮演任何角色。这里我们报告,468 - 530年HIP1构造是不稳定的,但矛盾的是在这个subfragment耐热结构单元存在。我们发现两个基本补丁的灵活的部分HIP1(集中在K474和K494)绑定的CLC至关重要。这些静电因素的一部分溶剂暴露疏水表面,我们以前认为是适合CLC。</pgydF4y2Ba>
<h4 id="results-and-discussion">2。结果与讨论</gydF4y2Bah4>
<h5 id="subsec2.1">2.1。卷曲螺旋段HIP1,包括网格蛋白轻链的决定因素在本质上是不稳定的</gydF4y2Bah5>
<p>绑定的CLC HIP1需要L486或R487 [<一个href="#B7">7</一个>](人类HIP1编号(005329年加入许多NP),在HIP12/1R守恒)。我们最近位于L486和R487灵活段HIP1(位置2,图<一个href="//www.newsama.com/journals/bri/2009/256124/fig1/" target="_blank">1</一个>),在S3中路径(<一个href="#B8">8</一个>]。在这里我们使用CD探针的稳定性HIP1 468 - 530(图的灰色地带<一个href="//www.newsama.com/journals/bri/2009/256124/fig1/" target="_blank">1</一个>Y468和R547)之间,S3的卷曲螺旋段路径。在<gydF4y2Basvg height="11.0375" id="M9" style="vertical-align:-0.17554pt;width:22.8125px;" version="1.1" viewbox="0 0 22.8125 11.0375" width="22.8125" xmlns="http://www.w3.org/2000/svg">
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</svg>由双CD概要文件是螺旋,表示最低在208和222海里(图<一个href="//www.newsama.com/journals/bri/2009/256124/fig2/" target="_blank">2</一个>)。强度的比例在222年和208海里可以用来区分从孤立的螺旋线螺旋线圈(<gydF4y2Basvg height="11.25" id="M10" style="vertical-align:-1.29163pt;width:15.175px;" version="1.1" viewbox="0 0 15.175 11.25" width="15.175" xmlns="http://www.w3.org/2000/svg">
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</svg>扫描,这表明HIP1 468 - 530包含一个热稳定元素。我们目前做的核磁共振实验,绘制出热不稳定的程度部分HIP1 468 - 530。</pgydF4y2Ba>
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<a href="http://www.pymol.org">http://www.pymol.org</一个>
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结构特点的HIP1网格蛋白光chain-binding网站。两个冷淡地分离基本氨基酸开放地区的HIP1调解网格蛋白轻链的绑定。组装模型生成使用PDB文件2 no2 [gydF4y2Ba)和2 qa7 (gydF4y2Ba]。为了清晰起见,我们只显示一部分第二HIP1螺旋(深灰色)Y468之前停止。黄色栏标记的位置暴露疏水性溶剂S3路径(gydF4y2Ba]。新的数据图gydF4y2Ba表明K474是绑定和暗示的强大的行列式S3路径DL之前开始gydF4y2BaKN地区。N和c终端HIP1晶体结构的标记N和c .沿着黄色数字1 - 3条马克氨基酸的位置,控制网格蛋白轻链的绑定(位置1:K474;职位2:L486和R487麦克弗森集团(gydF4y2Ba];和位置3:K494)。K474和K494 ~ 50gydF4y2Ba分开。R500 R508或K511(见箭头)不参与绑定,因此定义的边界光chain-binding网站。468 - 530年HIP1 subfragment用于CD研究图gydF4y2Ba跨越一个开放的区域HIP1卷曲螺旋2 no2和2 qa7晶体结构。使用PyMol HIP1模型被创建(gydF4y2Ba)。</gydF4y2Batable>
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HIP1 468 - 530 subfragment打开地区只在低温度是稳定的。平衡远紫外CD进行扫描gydF4y2Ba三角形(打开),(gydF4y2Ba打开钻石),(gydF4y2Ba交叉),(gydF4y2Ba开环)。222纳米的位置信号不随温度改变,但208海里信号转变为样本加热。222/208 nm比率从原始CD数据计算。因为信号低于200纳米是嘈杂的,我们没有试图得到螺旋从光盘内容数据。</gydF4y2Batable>
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<h5 id="subsec2.2">2.2。基本集中在K474和K494补丁HIP1对于绑定网格蛋白轻链至关重要</gydF4y2Bah5>
<p>我们调查如果开放地区的一系列基本的补丁可以参与绑定CLC。K494(见位置3,图<一个href="//www.newsama.com/journals/bri/2009/256124/fig1/" target="_blank">1</一个>)是在S3中路径(黄色栏图<一个href="//www.newsama.com/journals/bri/2009/256124/fig1/" target="_blank">1</一个>)之前描述的<一个href="#B8">8</一个>),紧随其后的是一群基本残留物(箭头在图所示<一个href="//www.newsama.com/journals/bri/2009/256124/fig1/" target="_blank">1</一个>)。K474(位置1)坐落在DL<gydF4y2Basvg height="14.5375" id="M21" style="vertical-align:-3.25793pt;width:54.162498px;" version="1.1" viewbox="0 0 54.162498 14.5375" width="54.162498" xmlns="http://www.w3.org/2000/svg">
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</svg>KN地区,靠近HIP1的一部分,介导杭丁顿蛋白相互作用蛋白1的绑定关联(HIPPI) [<一个href="#B9">9</一个>]。调查在S3中如果这些路径做出了贡献,我们5 GST-HIP1h (HIP1h aa370 - 644) 370 - 644突变体(K494A, K494E, R500E R508E, K511E)。我们进行了销售税的绑定下拉来评估6 hishub / 6 hislcb(网格蛋白的中心是中央第三(aa1074 - 1675) [<一个href="#B12">12</一个>)和6 hislcb氨基端histidine-tagged牛网格蛋白轻链与神经元插入b)。是重要研究6 hislcb绑定到6 hishub密切模仿HIP1如何与网格蛋白篮子在细胞相互作用。每个销售税下拉做了至少三次,使用新鲜分离出蛋白质和收取销售税珠子每一次。网格蛋白中心(氨基端组氨酸标记)是通过与商业组氨酸标签蛋白免疫印迹检测到抗体。LCb玷污了CON.1单克隆抗体和GST-HIP1h结构与商业销售税抗体可视化。anti-GST乐队的人物<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>和<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3 (b)</一个>表明GST-HIP1h水平平衡(加载控制)。消极的控制通道1 - 3在图<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>1 - 3在图和航线<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3 (b)</一个>从每个绑定的实验表明,消费税信号没有随机交互,但反映真实绑定事件。6的水平hishub / 6 hislcb被GST-HIP1h巷4图所示<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>和<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3 (b)</一个>,正如所料,需要绑定LCb(车道3和4的数据进行比较<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>和<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3 (b)</一个>)。我们没有删除组氨酸标签LCb用于净化因为控制实验表明,标签不干扰消费税下拉(数据没有显示)。绑定6 hishub / 6 hislcb明显阻塞时HIP1h K494取代了谷氨酸(K494E突变,莱茵5图<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>与丙氨酸)或(巷5,图<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3 (b)</一个>)。相比之下,我们看到当R500没有可检测的影响,K494正上方(见图<一个href="//www.newsama.com/journals/bri/2009/256124/fig1/" target="_blank">1</一个>位置),改变了谷氨酸(参见巷6,图<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>)或丙氨酸(数据未显示)。符合数据通道在图5和6<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>,K494E / R500E双突变体没有绑定6 hishub / 6 hislcb(图7巷<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>)。接下来我们评估一组基本残留接近S3的c端端路径(一个箭头在图所示<一个href="//www.newsama.com/journals/bri/2009/256124/fig1/" target="_blank">1</一个>)。车道在图8和9<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>表明R508E和K511E电荷反转突变体6 hishub / 6 hislcb类似于野生型控制(比较巷4车道在图8 - 9<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3(一个)</一个>)。我们从这些数据得出K494位于疏水性S3路径需要绑定CLC,但R500 R508, K511围绕这条路不参与。这表明CLC结合位点不超越S3的边界定义的路径由R500 R508, K511。</pgydF4y2Ba>
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<span class="caption-text">图3</gydF4y2Baspan>
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<table>
两种基本的补丁在HIP1绑定网格蛋白轻链的关键。西方墨迹图进行分析GST-pulldown实验。样本运行在相同的凝胶,但转移后,膜切开,涂抹与相应的抗体。牛网格蛋白重链(aa1074 - 1675,中心)和一个氨基端histidine-tag玷污了一个商业anti-His单克隆抗体(anti-His)。神经元的网格蛋白轻链b (LCb)氨基端histidine-tag,但是我们使用CON.1识别LCb。消费税仅和GST-HIP1h构造检测使用一个商业单克隆抗体销售税(Anti-GST)。面板:通道1和2:销售税仅负控制显示,没有特异性的相互作用6 hishub单独或6 hishub + 6 hislcb。6巷3:野生型GST-HIP1h下拉独自hishub(没有绑定LCb)表明,轻链的交互需求。巷4:积极控制,野生型与6 hishub / 6 hislcb GST-HIP1h。道5 - 9:下拉的HIP1h单引号和双突变体6 hishub / 6 hislcb。 Panel B: lanes 1–3 are controls as described in panel A. Lane 4: positive control, wild type GST-HIP1h with 6Hishub/6HisLCb. Lane 5: GST-HIP1h K494A with 6Hishub/6HisLCb. Lane 6: GST-HIP1h K474A with 6Hishub/6HisLCb.</table>
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<p>HIP1组装模型(2 no2加入到2 qa7)在图<一个href="//www.newsama.com/journals/bri/2009/256124/fig1/" target="_blank">1</一个>显示了积极的补丁以面向K474是相对于K494(由~ 50<gydF4y2Basvg height="15.15" id="M22" style="vertical-align:-0.0pt;width:11.4125px;" version="1.1" viewbox="0 0 11.4125 15.15" width="11.4125" xmlns="http://www.w3.org/2000/svg">
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</svg>)。我们的数据在图<一个href="//www.newsama.com/journals/bri/2009/256124/fig3/" target="_blank">3 (b)</一个>显示绑定的大幅下降6 hishub / 6 hislcb K474突变时丙氨酸(见4和6车道(K494A巷5进行比较)。这个结果表明,首次K474上游DL<gydF4y2Basvg height="14.5375" id="M23" style="vertical-align:-3.25793pt;width:54.162498px;" version="1.1" viewbox="0 0 54.162498 14.5375" width="54.162498" xmlns="http://www.w3.org/2000/svg">
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</svg>KN地区所需的绑定6 hishub / 6 hislcb表明CLC结合位点可能比之前认为的更广泛。我们预测,这里定义的两种基本的补丁在HIP1我们也存在于HIP12 / R因为K474 K494 (HIP1编号)在这种蛋白质都是守恒的。最后,我们看如果K474、R487 K494 HIP1从veterbrates守恒<gydF4y2Bai>亩骶</gydF4y2Bai>,<gydF4y2Bai>鼠形</gydF4y2Bai>,<gydF4y2Bai>非洲爪蟾蜍光滑的</gydF4y2Bai>,<gydF4y2Bai>黑腹果蝇</gydF4y2Bai>,在<gydF4y2Bai>sla2p</gydF4y2Bai>(HIP1同族体从<gydF4y2Bai>酿酒酵母</gydF4y2Bai>)和ZK370.3 (HIP1同族体<gydF4y2Bai>秀丽隐杆线虫</gydF4y2Bai>)。这一分析表明,K474保存所有的脊椎动物,但V<gydF4y2Bai>秀丽隐杆线虫</gydF4y2Bai>,我在<gydF4y2Bai>d .腹</gydF4y2Bai>在sla2p和M。R487守恒在所有的脊椎动物,但E<gydF4y2Bai>d .腹</gydF4y2Bai>和<gydF4y2Bai>sla2p</gydF4y2Bai>和T在<gydF4y2Bai>秀丽隐杆线虫</gydF4y2Bai>。K494守恒在所有的脊椎动物和无脊椎动物HIP1蛋白质,除了<gydF4y2Bai>sla2p</gydF4y2Bai>(K474(问<gydF4y2Bai>酿酒酵母</gydF4y2Bai>),R487 (T)和K494 (D))。自然突变<gydF4y2Bai>sla2p</gydF4y2Bai>,HIP1<gydF4y2Bai>d .腹</gydF4y2Bai>,ZK370.3<gydF4y2Bai>秀丽隐杆线虫</gydF4y2Bai>可能意味着HIP1绑定和HIP1同系物在这些生物网格蛋白轻链是不同的。</pgydF4y2Ba>
<h4 id="materials-and-methods">3所示。材料和方法</gydF4y2Bah4>
<h5 id="subsec3.1">3.1。材料</gydF4y2Bah5>
<p>特里同x - 100、Tween-20 beta-mercaptoethanol (<gydF4y2Bai>β</gydF4y2Bai>我),TRIZMA基地和BIS-TRIS来自Sigma-Aldrich(圣路易斯,密苏里州,美国)。磷酸氢二钠(N<gydF4y2Basvg height="11.225" id="M24" style="vertical-align:-3.13504pt;width:13.1625px;" version="1.1" viewbox="0 0 13.1625 11.225" width="13.1625" xmlns="http://www.w3.org/2000/svg">
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</svg>)从EMD化学品(美国新泽西州Gibbstown);仅有肉汤是EMD生物科学(火花,医学博士,美国);三羟甲基氨基甲烷(2-carboxyethyl)液磷化氢Sigma-Aldrich。Pfu涡轮是Stratagene(美国加州拉霍亚)和DNA引物来自集成技术(珊瑚镇,爱荷华州,美国)。皮尔斯Coomassie加上布拉德福德试剂盒购自费舍尔科学(美国汉诺威公园,生病)。色谱树脂、列和标准购买通用电气医疗集团(美国新泽西州皮斯卡塔韦)。CON.1抗体购自Covance(美国弗吉尼亚州坎伯兰);限制级凝血酶和抗体anti-His获得Novagen(美国加州拉霍亚)。Coomassie g - 250染色和Immun-Star化学发光设备来自Bio-Rad实验室(美国加州大力神)。</p>
<h5 id="subsec3.2">3.2。GST-HIP1h突变体和6 hislcb建设</gydF4y2Bah5>
<p>质粒编码最初的氨基端销售税标记HIP1h 370 - 644从麦克弗森集团是一个礼物。这项工作中使用的各种GST-HIP1h突变体创建使用QuikChange诱变协议(Stratagene)。证实了DNA测序序列(IMBI,印第安纳大学),然后变成了罗塞塔2 (DE3) pLysS细胞(Novagen)。标准克隆技术被用来插入神经元LCb DNA<gydF4y2Bai>p</gydF4y2Bai>6 hislcb ET15b生成。6 hislcb重组质粒的转化为BL21 (DE3) pLysS细胞表达。</pgydF4y2Ba>
<h5 id="subsec3.3">3.3。蛋白质超表达和纯化</gydF4y2Bah5>
<p>重组GST-HIP1h构造是增长的<gydF4y2Basvg height="11.0375" id="M26" style="vertical-align:-0.17555pt;width:30.625px;" version="1.1" viewbox="0 0 30.625 11.0375" width="30.625" xmlns="http://www.w3.org/2000/svg">
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</svg>(pH值7.3),140毫米氯化钠,氯化钾,2.7毫米的补充与德勤0.25毫升的1米,0.25毫升的蛋白酶抑制剂鸡尾酒(σ),和2毫升PMSF丙胺)(17.4毫克/毫升)。声波降解法后,2.5毫升的20% (v / v)特里同x - 100添加溶菌产物是旋转在室温下~ 30分钟。原油的细菌溶菌产物是000年12<gydF4y2Basvg height="10.725" id="M35" style="vertical-align:-2.7337pt;width:7.9375px;" version="1.1" viewbox="0 0 7.9375 10.725" width="7.9375" xmlns="http://www.w3.org/2000/svg">
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</svg>)10分钟。上层清液混合着~ 5毫升的谷胱甘肽琼脂糖4 b (Amersham)树脂悬浮在PBS。GST-HIP1h构造从50毫升的列筛选了3毫克/毫升L-glutathione(σ)(PBS) pH值8.0和透析一夜之间对相同的缓冲区。乳糜泻实验净化协议被修改以便我们能打通消费税在消费税列标签。细菌溶菌产物增加销售税珠子后,混合物在室温下是震撼了1 - 2小时。珠子是在500年<gydF4y2Basvg height="10.725" id="M37" style="vertical-align:-2.7337pt;width:7.9375px;" version="1.1" viewbox="0 0 7.9375 10.725" width="7.9375" xmlns="http://www.w3.org/2000/svg">
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</svg>5分钟,上层清液倒。潮湿的珠子被转移到一个列和110毫升1 x PBS慢慢洗,然后1单位凝血酶每毫克的蛋白质被稀释到3.5毫升1 x PBS和添加到列中。消化后,HIP1h Superdex 75列上构造进一步纯化产物(通用电气医疗集团)与1 x PBS(室温)。列分数是汇集和透析<gydF4y2Basvg height="11.0375" id="M39" style="vertical-align:-0.17554pt;width:22.8125px;" version="1.1" viewbox="0 0 22.8125 11.0375" width="22.8125" xmlns="http://www.w3.org/2000/svg">
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<p>网格蛋白中心建设是纯化如前所述<一个href="#B13">13</一个>)和6 hislcb在单个步骤使用镍亲和纯化树脂。在原油溶解产物添加到镍琼脂糖树脂和孵化<gydF4y2Basvg height="11.0375" id="M40" style="vertical-align:-0.17554pt;width:22.8125px;" version="1.1" viewbox="0 0 22.8125 11.0375" width="22.8125" xmlns="http://www.w3.org/2000/svg">
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</svg>10分钟,珠子轻轻旋转下来洗25毫升10毫米N<gydF4y2Basvg height="11.225" id="M41" style="vertical-align:-3.13504pt;width:13.1625px;" version="1.1" viewbox="0 0 13.1625 11.225" width="13.1625" xmlns="http://www.w3.org/2000/svg">
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</svg>,咪唑10毫米,0.5 M氯化钠,pH值7.4缓冲区(缓冲)。30毫升的珠子被洗缓冲区包含0.5米咪唑。6 hislcb列与缓冲区包含被筛选了205毫米咪唑。EDTA是添加到样本(1毫升0.5 EDTA每10毫升的蛋白质)和透析10毫米三羟甲基氨基甲烷、液pH值7.9一夜之间<gydF4y2Basvg height="11.0375" id="M43" style="vertical-align:-0.17554pt;width:22.8125px;" version="1.1" viewbox="0 0 22.8125 11.0375" width="22.8125" xmlns="http://www.w3.org/2000/svg">
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</svg>(透析缓冲区包括BME和蛋白酶抑制剂的鸡尾酒)。</pgydF4y2Ba>
<h5 id="subsec3.4">3.4。CD测量</gydF4y2Bah5>
<p>纯化HIP1 468 - 530稀释10毫米磷酸钾缓冲的pH值7.9至0.5毫克/毫升CD在不同的温度下测量。CD数据收集使用Jasco j - 175圆二色性与热控制样本细胞分光偏振计。是在第一张CD扫描<gydF4y2Basvg height="11.0375" id="M44" style="vertical-align:-0.17554pt;width:22.8125px;" version="1.1" viewbox="0 0 22.8125 11.0375" width="22.8125" xmlns="http://www.w3.org/2000/svg">
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</svg>然后改变了温度的温度图表示<一个href="//www.newsama.com/journals/bri/2009/256124/fig2/" target="_blank">2</一个>。样品被允许坐在每个表示温度前几分钟CD扫描。</pgydF4y2Ba>
<h5 id="subsec3.5">3.5。销售税拉下化验</gydF4y2Bah5>
<p>谷胱甘肽琼脂糖4 b树脂(通用电气医疗集团17-0756-01)与1毫升的PBS洗了三次。蛋白质浓度测定布拉德福德化验(皮尔斯23236)。等摩尔量的销售税和GST-Hip1h蛋白(<gydF4y2Basvg height="13.55" id="M45" style="vertical-align:-2.29482pt;width:36.5625px;" version="1.1" viewbox="0 0 36.5625 13.55" width="36.5625" xmlns="http://www.w3.org/2000/svg">
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</svg>)和1毫升的PBS被添加到树脂和孵化一个旋转平台上<gydF4y2Basvg height="11.0375" id="M46" style="vertical-align:-0.17554pt;width:22.8125px;" version="1.1" viewbox="0 0 22.8125 11.0375" width="22.8125" xmlns="http://www.w3.org/2000/svg">
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</g>
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</svg>1小时。释放的蛋白质被通过洗涤三次绑定缓冲(50毫米三,200毫米氯化钾,1毫米EDTA, 1% Triton x - 100, 50 mM咪唑,0.5毫克/毫升卵白蛋白,pH值8.0)。仅6 hislcb和中心结合在一个3:1比例和被允许孵化<gydF4y2Basvg height="11.0375" id="M47" style="vertical-align:-0.17554pt;width:22.8125px;" version="1.1" viewbox="0 0 22.8125 11.0375" width="22.8125" xmlns="http://www.w3.org/2000/svg">
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</svg>L销售税或GST-Hip1h绑定珠子,0.4 nmol中心单独或1.9 nmol中心6 hislcb复杂,375年<gydF4y2Basvg height="9.6750002" id="M49" style="vertical-align:-2.29482pt;width:9.6374998px;" version="1.1" viewbox="0 0 9.6374998 9.6750002" width="9.6374998" xmlns="http://www.w3.org/2000/svg">
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</svg>L室温绑定缓冲被添加到illustra MicroSpin列(通用电气医疗集团27-3565-01)和孵化在旋转<gydF4y2Basvg height="11.0375" id="M50" style="vertical-align:-0.17554pt;width:22.8125px;" version="1.1" viewbox="0 0 22.8125 11.0375" width="22.8125" xmlns="http://www.w3.org/2000/svg">
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</svg>1小时。珠子是洗6次为0.5毫升的绑定缓冲+ 16毫米咪唑和上层清液被离心分离。最后洗后,55<gydF4y2Basvg height="9.6750002" id="M51" style="vertical-align:-2.29482pt;width:9.6374998px;" version="1.1" viewbox="0 0 9.6374998 9.6750002" width="9.6374998" xmlns="http://www.w3.org/2000/svg">
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</svg>L 2 x SDS凝胶加载缓冲区添加珠子。自旋列被关闭放置在1.7毫升埃普多夫管,加热10分钟<gydF4y2Basvg height="11.1625" id="M52" style="vertical-align:-0.27588pt;width:46.25px;" version="1.1" viewbox="0 0 46.25 11.1625" width="46.25" xmlns="http://www.w3.org/2000/svg">
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</svg>,旋转收集样本。</pgydF4y2Ba>
<h5 id="subsec3.6">3.6。西方的屁股</gydF4y2Bah5>
<p>结合蛋白被解决通过sds - page和免疫印迹分析标准。转让后硝化纤维膜有朱红色污渍斑斑,减少分离蛋白质印迹。中心检测与anti-His单克隆抗体(Novagen 70796 - 3),销售税和发现了GST-Hip1h anti-GST单克隆抗体(Covance mms - 112 p)和6 hislcb检测与网格蛋白轻链单克隆抗体(CON.1) (Covance mms - 423 - p)。绑定与Immun-Star化学发光检测蛋白质检测系统(BioRad 170 - 5010)。</pgydF4y2Ba>
<h4 id="acknowledgments">确认</gydF4y2Bah4>
<p>我们感谢Sanjay Mishra生成一些GST-HIP1h构造用于这项工作,安娜Bonfitto蛋白质纯化和初始CD实验,和萨曼莎Perez-Miller创建6 hislcb构造。美国国立卫生研究院的资助(GM64387)是给第一作者为这项工作。</pgydF4y2Ba>
</div>
<div class="xml-content">
<h4 class="references" id="references">引用</gydF4y2Bah4>
<ol>
<li id="B1">一个。e . y . Engqvist-Goldstein m·m·凯塞尔v . s . Chopra, m·r·海登和d . g . Drubin”一个actin-binding Sla2 /杭丁顿蛋白相互作用蛋白1家族的蛋白质是一种新型组件clathrin-coated坑和囊泡的,”<gydF4y2Bai>《细胞生物学》杂志上</gydF4y2Bai>,卷147,不。7,1503 - 1518年,1999页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1083/jcb.147.7.1503">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=An%20actin-binding%20protein%20of%20the%20Sla2/Huntingtin%20interacting%20protein%201%20family%20is%20a%20novel%20component%20of%20clathrin-coated%20pits%20and%20vesicles&author=Å. E. Y. Engqvist-Goldstein&author=M. M. Kessels&author=V. S. Chopra&author=M. R. Hayden&author=&author=D. G. Drubin&publication_year=1999" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B2">s . k . Mishra n . r . Agostinelli t·j·布雷特Mizukami, t . s .罗斯和l·m·特劳布”网格蛋白——HIP1 AP-2-binding网站发现大会角色内吞作用的辅助蛋白质,”<gydF4y2Bai>《生物化学》杂志上</gydF4y2Bai>,卷276,不。49岁,46230 - 46236年,2001页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1074/jbc.M108177200">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Clathrin-%20and%20AP-2-binding%20sites%20in%20HIP1%20uncover%20a%20general%20assembly%20role%20for%20endocytic%20accessory%20proteins&author=S. K. Mishra&author=N. R. Agostinelli&author=T. J. Brett&author=I. Mizukami&author=T. S. Ross&author=&author=L. M. Traub&publication_year=2001" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B3">y太阳,m . Kaksonen d·t·马登r . Schekman和d . g . Drubin”交互Sla2p的尖刺外壳域与PtdIns actin-dependent (4、5) P2是重要的内吞作用的内化,”<gydF4y2Bai>细胞的分子生物学</gydF4y2Bai>,16卷,不。2、717 - 730年,2005页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1091/mbc.E04-08-0740">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Interaction%20of%20Sla2p's%20ANTH%20domain%20with%20PtdIns(4,5)P2%20is%20important%20for%20actin-dependent%20endocytic%20internalization&author=Y. Sun&author=M. Kaksonen&author=D. T. Madden&author=R. Schekman&author=&author=D. G. Drubin&publication_year=2005" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B4">t . m . Newpher F.-Z。m . i Geli Idrissi, s . k .雷蒙”小说网格蛋白轻链的功能在促进内吞作用的囊泡的形成,“<gydF4y2Bai>细胞的分子生物学</gydF4y2Bai>,17卷,不。10日,4343 - 4352年,2006页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1091/mbc.E06-07-0606">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Novel%20function%20of%20clathrin%20light%20chain%20in%20promoting%20endocytic%20vesicle%20formation&author=T. M. Newpher&author=F.-Z. Idrissi&author=M. I. Geli&author=&author=S. K. Lemmon&publication_year=2006" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B5">t·j·布雷特·l·m·特劳布·d·h·弗里蒙特,“辅助蛋白质招聘主题clathrin-mediated内吞作用,”<gydF4y2Bai>结构</gydF4y2Bai>,10卷,不。6,797 - 809年,2002页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1016/S0969-2126(02)00784-0">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Accessory%20protein%20recruitment%20motifs%20in%20clathrin-mediated%20endocytosis&author=T. J. Brett&author=L. M. Traub&author=&author=D. H. Fremont&publication_year=2002" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B6">t·j·布雷特诉Legendre-Guillemin·s·麦克弗森和d·h·弗里蒙特”结构的定义F-actin-binding茅草HIP1R域,“<gydF4y2Bai>自然结构和分子生物学</gydF4y2Bai>,13卷,不。2、121 - 130年,2006页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1038/nsmb1043">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Structural%20definition%20of%20the%20F-actin-binding%20THATCH%20domain%20from%20HIP1R&author=T. J. Brett&author=V. Legendre-Guillemin&author=P. S. McPherson&author=&author=D. H. Fremont&publication_year=2006" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B7">诉Legendre-Guillemin麦茨勒·m·肯尼迪。勒麦尔et al .,“杭丁顿蛋白相互作用蛋白1 (HIP1)调节网格蛋白组装通过直接绑定到监管区域的网格蛋白轻链,”<gydF4y2Bai>《生物化学》杂志上</gydF4y2Bai>,卷280,不。7,6101 - 6108年,2005页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1074/jbc.M408430200">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Huntingtin%20interacting%20protein%201%20(HIP1)%20regulates%20clathrin%20assembly%20through%20direct%20binding%20to%20the%20regulatory%20region%20of%20the%20clathrin%20light%20chain&author=V. Legendre-Guillemin&author=M. Metzler&author=J.-F. Lemaire et al.&publication_year=2005" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B8">j . a .状态s Mishra s .头盔和j . Nix“晶体结构的2.8 DLLRKN-containing卷曲螺旋域Huntingtin-interacting蛋白1 (HIP1)揭示了一个表面适合网格蛋白轻链绑定,”<gydF4y2Bai>分子生物学杂志</gydF4y2Bai>,卷367,不。1、地位,2007页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1016/j.jmb.2006.12.052">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Crystal%20structure%20at%202.8Å%20of%20the%20DLLRKN-containing%20coiled-coil%20domain%20of%20Huntingtin-interacting%20protein%201%20(HIP1)%20reveals%20a%20surface%20suitable%20for%20clathrin%20light%20chain%20binding&author=J. A. Ybe&author=S. Mishra&author=S. Helms&author=&author=J. Nix&publication_year=2007" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B9">问:妞妞和j . a .状态”,晶体结构在2.8 Huntingtin-interacting蛋白1 (HIP1)卷曲螺旋域显示一个带电表面适合HIP1蛋白质关联(HIPPI)”<gydF4y2Bai>分子生物学杂志</gydF4y2Bai>,卷375,不。5,1197 - 1205年,2008页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1016/j.jmb.2007.11.036">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Crystal%20structure%20at%202.8Å%20of%20Huntingtin-interacting%20protein%201%20(HIP1)%20coiled-coil%20domain%20reveals%20a%20charged%20surface%20suitable%20for%20HIP1%20protein%20interactor%20(HIPPI)&author=Q. Niu &author=J. A. Ybe&publication_year=2008" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B10">郑胜耀m·刘,a·k·希夫,r·s·霍奇斯”合成一个模型的定义二级和四级结构的蛋白质。链长对稳定的影响和形成两股<gydF4y2Bamath id="M53" xmlns="http://www.w3.org/1998/Math/MathML">
<mrow>
<mi>
α</gydF4y2Bami>
</mrow>
</math>螺旋coiled-coils。”<gydF4y2Bai>《生物化学》杂志上</gydF4y2Bai>,卷259,不。21日,第13261 - 13253页,1984年。<gydF4y2Baspan class="reflinks">视图:<一个href="https://scholar.google.com/scholar_lookup?title=Synthesis%20of%20a%20model%20protein%20of%20defined%20secondary%20and%20quaternary%20structure.%20Effect%20of%20chain%20length%20on%20the%20stabilization%20and%20formation%20of%20two-stranded%20a-helical%20coiled-coils&author=S. Y. M. Lau&author=A. K. Taneja&author=&author=R. S. Hodges&publication_year=1984" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B11">c .麦克纳马拉A . s . Zinkernagel p . Macheboeuf m·w·坎宁安诉Nizet和p . Ghosh“卷曲螺旋不规则和不稳定A群链球菌M1所需毒性,”<gydF4y2Bai>科学</gydF4y2Bai>,卷319,不。5868年,第1408 - 1405页,2008年。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1126/science.1154470">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Coiled-coil%20irregularities%20and%20instabilities%20in%20group%20A%20Streptococcus%20M1%20are%20required%20for%20virulence&author=C. McNamara&author=A. S. Zinkernagel&author=P. Macheboeuf&author=M. W. Cunningham&author=V. Nizet&author=&author=P. Ghosh&publication_year=2008" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B12">工程学系。刘,m . l . Wong c . s . Craik和f·m·布罗斯基”的监管网格蛋白组装和三聚使用重组三曲臂图定义的中心,“<gydF4y2Bai>细胞</gydF4y2Bai>,卷83,不。2、257 - 267年,1995页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1016/0092-8674(95)90167-1">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Regulation%20of%20clathrin%20assembly%20and%20trimerization%20defined%20using%20recombinant%20triskelion%20hubs&author=S.-H. Liu&author=M. L. Wong&author=C. S. Craik&author=&author=F. M. Brodsky&publication_year=1995" target="_blank">谷歌学术搜索</一个></span></li>
<li id="B13">j . a .状态s Perez-Miller问:妞妞,d·a·科茨·m·w·德拉泽在m·e·克莱格,“轻链c端地区强化网格蛋白重链的稳定性三,”<gydF4y2Bai>交通</gydF4y2Bai>,8卷,不。8,1101 - 1110年,2007页。<gydF4y2Baspan class="reflinks">视图:<一个href="https://doi.org/10.1111/j.1600-0854.2007.00597.x">出版商的网站</一个><span class="sep">|</gydF4y2Baspan><a href="https://scholar.google.com/scholar_lookup?title=Light%20chain%20C-terminal%20region%20reinforces%20the%20stability%20of%20clathrin%20heavy%20chain%20trimers&author=J. A. Ybe&author=S. Perez-Miller&author=Q. Niu&author=D. A. Coates&author=M. W. Drazer&author=&author=M. E. Clegg&publication_year=2007" target="_blank">谷歌学术搜索</一个></span></li>
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<p>版权©2009乔尔·a·状态等。这是一个开放的分布式下文章<一个rel="license" href="http://creativecommons.org/licenses/by/3.0/">知识共享归属许可</一个>,它允许无限制的使用、分配和复制在任何媒介,提供最初的工作是正确引用。</pgydF4y2Ba>
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的观点<gydF4y2Baspan>769年</gydF4y2Baspan>
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下载<gydF4y2Baspan>966年</gydF4y2Baspan>
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<span class="__dimensions_badge_embed__" data-doi="10.1155/2009/256124" data-style="small_circle"></span>引用</gydF4y2Badiv>
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