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Noncollagenous region of the streptococcal collagen‐like protein is a trimerization domain that supports refolding of adjacent homologous and heterologous collagenous domains
Author(s) -
Yu Zhuoxin,
Mirochnitchenko Oleg,
Xu Chunying,
Yoshizumi Ayumi,
Brodsky Barbara,
Inouye Masayori
Publication year - 2010
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.356
Subject(s) - heterologous , homologous chromosome , domain (mathematical analysis) , protein superfamily , chemistry , biophysics , microbiology and biotechnology , crystallography , biology , biochemistry , mathematics , gene , mathematical analysis
Proper folding of the (Gly‐Xaa‐Yaa) n sequence of animal collagens requires adjacent N‐ or C‐terminal noncollagenous trimerization domains which often contain coiled‐coil or beta sheet structure. Collagen‐like proteins have been found recently in a number of bacteria, but little is known about their folding mechanism. The Scl2 collagen‐like protein from S treptococcus pyogenes has an N‐terminal globular domain, designated V sp , adjacent to its triple‐helix domain. The V sp domain is required for proper refolding of the Scl2 protein in vitro . Here, recombinant V sp domain alone is shown to form trimers with a significant α‐helix content and to have a thermal stability of T m = 45°C. Examination of a new construct shows that the V sp domain facilitates efficient in vitro refolding only when it is located N‐terminal to the triple‐helix domain but not when C‐terminal to the triple‐helix domain. Fusion of the V sp domain N‐terminal to a heterologous (Gly‐Xaa‐Yaa) n sequence from C lostridium perfringens led to correct folding and refolding of this triple‐helix, which was unable to fold into a triple‐helical, soluble protein on its own. These results suggest that placement of a functional trimerization module adjacent to a heterologous Gly‐Xaa‐Yaa repeating sequence can lead to proper folding in some cases but also shows specificity in the relative location of the trimerization and triple‐helix domains. This information about their modular nature can be used in the production of novel types of bacterial collagen for biomaterial applications.