title: Preparation and characterization of monomers to tetramers of a collagen-like domain from Streptococcus pyogenes.
abstract: The collagen like domain Scl2 from Streptococcus pyogenes has been proposed as a potential biomedical material. It is non-cytotoxic and non-immunogenic and can be prepared in good yield in fermentation. The Scl2 collagen domain is about a quarter of the length, 234 residues, of the main collagen type, mammalian type I collagen (1014 residues) that is currently used in biomedical devices. In the present study we have made constructs comprising 1 to 4 copies of the Scl2 collagen domain, plus these same constructs with a CysCys sequence at the C-terminal, analogous to that found in mammalian type III collagens. The yields of these constructs were examined from 2 L fermentation studies. The yields of both series declined with increasing size. Circular dichroism showed that the addition of further collagen domains did not lead to a change in the melting temperature compared to the monomer domain. Addition of the CysCys sequence led to a small additional stabilization of about 2-3°C for the monomer construct when the folding (V) domain was present.
authors.affiliation: a CSIRO Manufacturing Flagship ; Clayton , Australia.
authors.forename: Yong Y
authors.initials: YY
authors.lastname: Peng
authors.forename: Violet
authors.initials: V
authors.lastname: Stoichevska
authors.forename: Linda
authors.initials: L
authors.lastname: Howell
authors.forename: Soren
authors.initials: S
authors.lastname: Madsen
authors.forename: Jerome A
authors.initials: JA
authors.lastname: Werkmeister
authors.forename: Geoff J
authors.initials: GJ
authors.lastname: Dumsday
authors.forename: John A M
authors.initials: JA
authors.lastname: Ramshaw
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D003094:Collagen / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013297:Streptococcus pyogenes / Q000378:metabolism*; D044366:Transition Temperature
chemical_list: D001426:Bacterial Proteins; D003094:Collagen
keywords: CL, collagenous domain of the S. pyogenes collagen-like protein; V, non-collagenous N-terminal domain of the S. pyogenes collagen-like protein; VCL, full collagen-like protein structure; collagen; fermentation; melting temperature; protein polymers; recombinant