Solution NMR structures of IgG binding domains with artificially evolved high levels of sequence identity but different folds

被引:24
作者
He, YN [1 ]
Yeh, DC [1 ]
Alexander, P [1 ]
Bryan, PN [1 ]
Orban, J [1 ]
机构
[1] Univ Maryland, Biotechnol Inst, Ctr Adv Res Biotechnol, Rockville, MD 20850 USA
关键词
D O I
10.1021/bi051232j
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 describe here the solution NMR structures of two IgG binding domains with highly homologous sequences but different three-dimensional structures. The proteins, G311 and A219, are derived from the IgG binding domains of their wild-type counterparts, protein G and protein A, respectively. Through a series of site-directed mutations and phage display selections, the sequences of G311 and A219 were designed to converge to a point of high-level sequence identity while keeping their respective wild-type tertiary folds. Structures of both artificially evolved sequences were determined by NMR spectroscopy. The main chain fold of G311 can be superimposed on the wild-type alpha/beta protein G structure with a backbone rmsd of 1.4 angstrom, and the A219 structure can be overlaid on the wild-type three-alpha-helix protein A fold also with a backbone rmsd of 1.4 angstrom. The structure of G311, in particular, accommodates a large number of mutational changes without undergoing a change in the overall fold of the main chain. The structural differences are maintained despite a high level (59%) of sequence identity. These proteins serve as starting points for further experiments that will probe basic concepts of protein folding and conformational switching.
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收藏
页码:14055 / 14061
页数:7
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