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首页> 外文期刊>Journal of Molecular Biology >Rolling adhesion of alpha(L) I domain mutants decorrelated from binding affinity
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Rolling adhesion of alpha(L) I domain mutants decorrelated from binding affinity

机译：与结合亲和力相关的α（L）I结构域突变体的滚动粘附

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Activated lymphocyte function-associated antigen-1 (LFA-1, alpha(L)beta(2) integrin) found on leukocytes facilitates firm adhesion to endothelial cell layers by binding to intercellular adhesion molecule-1 (ICAM-1), which is upregulated on endothelial cells at sites of inflammation. Recent work has shown that LFA-1 in a pre-activation, low-affinity state may also be involved in the initial tethering and rolling phase of the adhesion cascade. The inserted (1) domain of LFA-1 contains the ligand-binding epitope of the molecule, and a conformational change in this region during activation increases ligand affinity. We have displayed wild-type I domain on the surface of yeast and validated expression using I domain specific antibodies and flow cytometry. Surface display of I domain supports yeast rolling on ICAM-1-coated surfaces wider shear flow. Expression of a locked open, high-affinity I domain mutant supports firm adhesion of yeast, while yeast displaying intermediate-affinity I domain mutants exhibit a range of rolling phenotypes. We find that rolling behavior for these mutants fails to correlate with ligand binding affinity. These results indicate that unstressed binding affinity is not the only molecular property that determines adhesive behavior under shear flow. (c) 2006 Elsevier Ltd. All rights reserved.
机译：在白细胞上发现活化的淋巴细胞功能相关抗原1（LFA-1，alpha（L）beta（2）整合素）通过与细胞间黏附分子1（ICAM-1）结合而促进了对内皮细胞层的牢固黏附在炎症部位的内皮细胞上。最近的工作表明，处于激活前，低亲和力状态的LFA-1也可能参与了粘附级联的初始束缚和滚动阶段。 LFA-1的插入的（1）域包含分子的配体结合表位，并且在激活过程中此区域的构象变化会增加配体亲和力。我们已经在酵母表面展示了野生型I结构域，并使用I结构域特异性抗体和流式细胞仪验证了表达。 I结构域的表面展示支持酵母在ICAM-1包被的表面上滚动，产生更大的剪切流。锁定的开放的高亲和力I结构域突变体的表达支持酵母的牢固粘附，而展示中间亲和力I结构域突变体的酵母则表现出一系列滚动表型。我们发现这些突变体的滚动行为无法与配体结合亲和力相关。这些结果表明，不受应力的结合亲和力不是决定剪切流下粘合行为的唯一分子性质。 （c）2006 Elsevier Ltd.保留所有权利。

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