| Autor: |
Zhuoxin Yu, Oleg Mirochnitchenko, Ayumi Yoshizumi, Barbara Brodsky, Chunying Xu, Masayori Inouye |
| Rok vydání: |
2010 |
| Předmět: |
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| Zdroj: |
Protein Science. 19:775-785 |
| ISSN: |
0961-8368 |
| DOI: |
10.1002/pro.356 |
| Popis: |
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 Streptococcus pyogenes has an N-terminal globular domain, designated Vsp, adjacent to its triple-helix domain. The Vsp domain is required for proper refolding of the Scl2 protein in vitro. Here, recombinant Vsp domain alone is shown to form trimers with a significant α-helix content and to have a thermal stability of Tm = 45°C. Examination of a new construct shows that the Vsp 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 Vsp domain N-terminal to a heterologous (Gly-Xaa-Yaa)n sequence from Clostridium 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. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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