The unique properties of the collagen triple-helix motif include its molecular hydrodynamic properties, extensive hydration, ability to bind diverse ligands, and the capacity self-associate to form fibrils and other higher order structures. These distinctive features have been exploited by nature to fill a wide range of structural and functional niches, and by scientists to design numerous products important for biomaterial and biomedical uses. Because of problems and concerns with the use of extracted collagens, attention has shifted to production of recombinant animal collagens, but efficient production has been slowed down addressing issues of post-translational modification. Rather than focus on specific types of animal collagens, we propose to focus on the intrinsic properties of the motif, using a bacterial triple-helix module which has high stability with no post-translational modifications. Stable triple-helix modules based on bacterial """"""""collagen-like"""""""" sequences will be utilized to form triple-helical polymers in a high yield E. coli cold shock expression system. Repeating modules will be used to attain lengths comparable to that of animal fibril forming collagens, and we hypothesize that these triple-helical proteins will self-assemble to form fibrillar structures. Defined human collagen sequences involved in known interactions will be inserted between adjacent bacterial collagen modules to produce chimeras with functional binding sites. Introduction of basement membrane type breaks in the repeating (Gly-X-Y)n sequence between bacterial triple-helix modules will used to create a flexible or kinked type structure that more closely models basement membrane collagen networks and may provide a substrate for stem cell cultures. This novel approach plans to capture the flexibility and advantages of the collagen triple-helix motif while retaining the simplicity of a bacterial expression system. The products of this system will be ideal for manipulations and applications as scaffolds in tissue engineering. Lay Summary: A novel approach to gain efficient production of repeating bacterial collagen-like sequences plans to capture the flexibility and advantages of the collagen triple-helix motrf while retaining the simplicity of a bacterial expression system. The products of this system will be ideal for manipulations and applications as scaffolds in tissue engineering. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21EB007198-02
Application #
7296100
Study Section
Musculoskeletal Tissue Engineering Study Section (MTE)
Program Officer
Hunziker, Rosemarie
Project Start
2006-09-29
Project End
2009-08-31
Budget Start
2007-09-01
Budget End
2009-08-31
Support Year
2
Fiscal Year
2007
Total Cost
$264,233
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Biochemistry
Type
Schools of Medicine
DUNS #
617022384
City
Piscataway
State
NJ
Country
United States
Zip Code
08854
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Yu, Zhuoxin; Mirochnitchenko, Oleg; Xu, Chunying et al. (2010) Noncollagenous region of the streptococcal collagen-like protein is a trimerization domain that supports refolding of adjacent homologous and heterologous collagenous domains. Protein Sci 19:775-85
Xu, Chunying; Yu, Zhuoxin; Inouye, Masayori et al. (2010) Expanding the family of collagen proteins: recombinant bacterial collagens of varying composition form triple-helices of similar stability. Biomacromolecules 11:348-56
Peng, Yong Y; Yoshizumi, Ayumi; Danon, Stephen J et al. (2010) A Streptococcus pyogenes derived collagen-like protein as a non-cytotoxic and non-immunogenic cross-linkable biomaterial. Biomaterials 31:2755-61
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Mohs, Angela; Silva, Teresita; Yoshida, Takeshi et al. (2007) Mechanism of stabilization of a bacterial collagen triple helix in the absence of hydroxyproline. J Biol Chem 282:29757-65