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A phage display system for studying the sequence determinants of protein folding
Author(s) -
Gu Hongdi,
Yi Qian,
Bray Susan T.,
Riddle David S.,
Baker David,
Shiau Andrew K.
Publication year - 1995
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560040609
Subject(s) - computational biology , phage display , protein design , protein folding , selection (genetic algorithm) , genetics , protein engineering , biology , sequence (biology) , population , folding (dsp implementation) , point mutation , protein structure , mutation , computer science , biochemistry , antibody , gene , sociology , enzyme , demography , artificial intelligence , electrical engineering , engineering
We have developed a phage display system that provides a means to select variants of the IgG binding domain of peptostreptococcal protein L that fold from large combinatorial libraries. The premise underlying the selection scheme is that binding of protein L to IgG requires that the protein be properly folded. Using a combination of molecular biological and biophysical methods, we show that this assumption is valid. First, the phage selection procedure strongly selects against a point mutation in protein L that disrupts folding but is not in the IgG binding interface. Second, variants recovered from a library in which the first third of protein L was randomized are properly folded. The degree of sequence variation in the selected population is striking: the variants have as many as nine substitutions in the 14 residues that were mutagenized. The approach provides a selection for “foldedness” that is potentially applicable to any small binding protein.