Computationally designed transmembrane α-helical peptides (CHAMP) have been used to compete for helix-helix interactions within the membrane, enabling the ability to probe the activation of the integrins αIIbβ3 and αvβ3. Here, this method is extended towards the design of CHAMP peptides that inhibit the association of the α5β1 transmembrane (TM) domains, targeting the Ala-X3-Gly motif within α5. Our previous design algorithm was performed alongside a new workflow implemented within the widely used Rosetta molecular modeling suite. Peptides from each computational approach activated integrin α5β1 but not αVβ3 in human endothelial cells. Two CHAMP peptides were shown to directly associate with an α5 TM domain peptide in detergent micelles to a similar degree as a β1 TM peptide does. By solution-state nuclear magnetic resonance, one of these CHAMP peptides was shown to bind primarily the integrin β1 TM domain, which itself has a Gly-X3-Gly motif. The second peptide associated modestly with both α5 and β1 constructs, with slight preference for α5. Although the design goal was not fully realized, this work characterizes novel CHAMP peptides activating α5β1 that can serve as useful reagents for probing integrin biology.

中文翻译：

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从头设计了激活α5β1整联蛋白的跨膜肽。

计算设计的跨膜α-螺旋肽（CHAMP）已用于竞争膜内的螺旋-螺旋相互作用，从而能够探测整合素αIIbβ3和αvβ3的激活。在此，此方法扩展到了CHAMP肽的设计，该肽可抑制α5β1跨膜（TM）域的结合，并靶向α5中的Ala-X3-Gly基序。我们先前的设计算法与在广泛使用的Rosetta分子建模套件中实施的新工作流程一起执行。来自每种计算方法的肽在人内皮细胞中激活整联蛋白α5β1，但不激活αVβ3。已显示两个CHAMP肽与去污剂胶束中的α5TM结构域肽直接缔合，其程度与β1TM肽相似。通过溶液态核磁共振，这些CHAMP肽之一显示主要结合整联蛋白β1TM结构域，该结构域本身具有Gly-X3-Gly基序。第二肽适度地与α5和β1构建体缔合，略微偏爱α5。尽管设计目标尚未完全实现，但这项工作还是表征了激活α5β1的新型CHAMP肽，该肽可用作探测整联蛋白生物学的有用试剂。