展示二硫键约束微蛋白的M13噬菌体

M13 bacteriophage displaying disulfide-constrained microproteins.

作者信息

McLafferty M A, Kent R B, Ladner R C, Markland W

机构信息

Protein Engineering Corporation, Cambridge, MA 02138.

出版信息

Gene. 1993 Jun 15;128(1):29-36. doi: 10.1016/0378-1119(93)90149-w.

DOI:10.1016/0378-1119(93)90149-w

PMID:8508957

Abstract

A display-phage library (TN2), displaying an 18-residue peptide fused to coat protein III, represents a collection of up to 8.55 x 10(6) peptides encoded by only 1.68 x 10(7) DNA sequences. Each displayed peptide has two fixed cysteine residues (allowing disulfide formation) and six variegated residues, four between the cysteines and one either side of the cysteines. Screening this library against streptavidin (Sv) and the anti-beta-endorphin monoclonal antibody, 3-E7, yielded phage displaying disulfide-constrained microproteins with sequences similar to those published for the linear-peptide display phage. Analysis of selected clones indicated that a disulfide bond is required for high-affinity binding to each of the target proteins. The microproteins selected for binding to Sv and 3-E7 show more stringent sequence specificity than do linear peptides selected for binding to the same targets.

摘要

一个展示噬菌体文库（TN2），其展示与外壳蛋白III融合的18个残基的肽，代表了由仅1.68×10⁷个DNA序列编码的多达8.55×10⁶个肽的集合。每个展示的肽有两个固定的半胱氨酸残基（允许形成二硫键）和六个可变残基，四个在半胱氨酸之间，一个在半胱氨酸的两侧。用抗生物素蛋白（Sv）和抗β-内啡肽单克隆抗体3-E7筛选该文库，得到了展示二硫键约束微蛋白的噬菌体，其序列与已发表的线性肽展示噬菌体的序列相似。对选定克隆的分析表明，二硫键是与每种靶蛋白高亲和力结合所必需的。选择与Sv和3-E7结合的微蛋白比选择与相同靶标结合的线性肽表现出更严格的序列特异性。