title: Design, expression and characterization of collagen-like proteins based on the cell adhesive and crosslinking sequences derived from native collagens.
abstract: Two recombinant collagen-like proteins consisting of cell adhesion domains derived from native type I collagen were designed and synthesized by a genetic engineering method. The cross-linking sequence, GPPGPCCGGG, derived from collagen III was used to promote triple helix formation through the disulfide bonds formed among three chains by flanking the peptide at the C-terminal of the collagen-like proteins. SDS-PAGE and western-blotting data suggested possibility of the formation of a triple helix structure for both recombinant proteins. CD spectra and thermal stability analyses indicated that the triple-helix structure in the collagen-like proteins was pH-dependent and stabilized under acidic environmental condition. Moreover, the collagen-like protein flanked with the cross-linking sequence at the C-terminal showed the most stable triple-helical conformation under acidic conditions.
authors.affiliation: Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan.
authors.forename: Juming
authors.initials: J
authors.lastname: Yao
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yanagisawa
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Asakura
mesh_terms: D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D002942:Circular Dichroism; D003094:Collagen / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics*; D005818:Genetic Engineering; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry*; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics*; D011994:Recombinant Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D013696:Temperature
chemical_list: D009841:Oligonucleotides; D010446:Peptide Fragments; D011506:Proteins; D011994:Recombinant Proteins; D003094:Collagen