In vivo biosynthesis of an Ala-scan library based on the cyclic peptide SFTI-1

被引：53

作者：

Austin, Jeffrey ^{[2
]}

Kimura, Richard H. ^{[2
]}

Woo, Youn-Hi ^{[2
]}

Camarero, Julio A. ^{[1
,2
]}

机构：

[1] Univ So Calif, Sch Pharm, Dept Pharmaceut Sci & Pharmacol, Los Angeles, CA 90033 USA

[2] Lawrence Livermore Natl Lab, Livermore, CA 94550 USA

来源：

AMINO ACIDS | 2010年 / 38卷 / 05期

关键词：

Bowman-Birk inhibitor; Trypsin inhibitor; Backbone cyclized peptides; Genetically encoded libraries; Protein splicing; SUNFLOWER TRYPSIN INHIBITOR-1; EXPRESSED PROTEIN LIGATION; MACROCYCLIC PEPTIDES; CIRCULAR PROTEINS; CHEMICAL LIGATION; SPLIT INTEIN; DRUG DESIGN; DNAE GENE; CYCLIZATION; CYCLOTIDES;

D O I：

10.1007/s00726-009-0338-4

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We present the in vivo biosynthesis of wild-type sunflower trypsin inhibitor 1 (SFTI-1) inside E. coli cells using an intramolecular native chemical ligation in combination with a modified protein splicing unit. SFTI-1 is a small backbone cyclized polypeptide with a single disulfide bridge. A small library containing multiple Ala mutants was also biosynthesized and its activity was assayed using a trypsin-binding assay. This study clearly demonstrates the exciting possibility of generating large cyclic peptide libraries in live E. coli cells, and is a critical first step for developing in vivo screening and directed evolution technologies using the cyclic peptide SFTI-1 as a molecular scaffold.

引用

页码：1313 / 1322

页数：10

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