Zou Yekui, Yin Jun
Department of Chemistry, The University of Chicago, 929 E. 57th Street, GCIS, E505A, Chicago, IL 60637, USA.
Bioorg Med Chem Lett. 2008 Oct 15;18(20):5664-7. doi: 10.1016/j.bmcl.2008.08.085. Epub 2008 Aug 28.
To engineer the substrate specificities of nonribosomal peptide synthetases (NRPS), we developed a method to display NRPS modules on M13 phages and select catalytically active adenylation (A) domains that would load azide functionalized substrate analogs to the neighboring peptidyl carrier protein (PCP) domains. Biotin conjugated difluorinated cyclooctyne was used for copper free cycloaddition with an azide substituted substrate attached to PCP. Biotin-labeled phages were selected by binding to streptavidin.
为了改造非核糖体肽合成酶（NRPS）的底物特异性，我们开发了一种方法，将NRPS模块展示在M13噬菌体上，并选择具有催化活性的腺苷化（A）结构域，该结构域可将叠氮化物功能化的底物类似物加载到相邻的肽基载体蛋白（PCP）结构域上。生物素偶联的二氟环辛炔用于与连接到PCP上的叠氮化物取代底物进行无铜环加成反应。通过与链霉亲和素结合来选择生物素标记的噬菌体。
