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首页> 外文期刊>Journal of Molecular Biology >Dissection of bacteriophage lambda site-specific recombination using synthetic peptide combinatorial libraries.
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Dissection of bacteriophage lambda site-specific recombination using synthetic peptide combinatorial libraries.

机译：使用合成肽组合文库解剖噬菌体λ位点特异性重组。

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摘要

A wide variety of tools have been used to dissect biochemical pathways, inhibitors being chief among them. Combinatorial approaches have made the search for inhibitors much more efficient. We have applied such an approach to identify hexapeptides which inhibit different steps in a site-specific recombination reaction mediated by the bacteriophage lambda integrase protein. Integrase's mechanism is still incompletely understood, in large part because several pathway intermediates remain hard to isolate. Integrase-catalyzed recombination is very efficient, but if blocked, it is highly reversible to substrates; this combination makes some intermediates exceedingly transient. We have used synthetic peptide combinatorial libraries to screen for hexapeptides that affect the recombination pathway at different stages, and have identified two families of peptides: one probably blocks DNA cleavage, the other may stabilize the Holliday junction intermediates. These peptides do not resemble parts of integrase or any of the other helper functions in the pathway. The deconvolution of hexapeptide libraries based both on inhibition of an enzymatic reaction as well as on accumulation of reaction intermediates is a novel approach to finding useful tools for dissecting a biochemical pathway. Copyright 2000 Academic Press.
机译：已经使用了各种各样的工具来剖析生化途径，其中抑制剂是主要的。组合方法使寻找抑制剂的效率大大提高。我们已经应用了这种方法来鉴定六肽，该六肽抑制由噬菌体λ整合酶蛋白介导的位点特异性重组反应中的不同步骤。对Integrase的机制仍未完全了解，在很大程度上是因为几个途径的中间体仍然难以分离。整合酶催化的重组非常有效，但如果被封闭，则对底物高度可逆;这种结合使某些中间体非常短暂。我们已经使用合成的肽组合文库筛选了在不同阶段影响重组途径的六肽，并鉴定了两个肽家族：一个可能阻断DNA裂解，另一个可能稳定霍利迪连接中间体。这些肽与该途径中整合酶的部分或任何其他辅助功能不相似。基于对酶促反应的抑制以及反应中间体的积累，对六肽文库的反卷积是一种发现用于剖析生化途径的有用工具的新颖方法。版权所有2000学术出版社。

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