Production and characterization of a silk-like hybrid protein, based on the polyalanine region of Samia cynthia ricini silk fibroin and a cell adhesive region derived from fibronectin.
There are a variety of silkworms and silk fibroins produced by them. Silks have many inherent suitable properties for biomaterials. In this paper, a novel silk-like hybrid protein, [DGG(A)(12)GGAASTGRGDSPAAS](5), which consists of polyalanine region of silk fibroin from a wild silkworm, Samia cynthia ricini, and cell adhesive region including Arg-Gly-Asp (RGD) sequence, derived from fibronectin, was designed and produced. The genes encoding the hybrid protein were constructed and expressed in Escherichia coli. The main conformation of the polyalanine region, that is, either alpha-helix or beta-sheet, could be easily controlled by treatment with different acidic solvents, trifluoroacetic acid or formic acid, respectively. This structural change was monitored with 13C CP/MAS NMR. Higher cell adhesive and growth activities of the hybrid protein compared with those of collagen were obtained.
Asakura
Tetsuo
T
Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan. asakura@cc.tuat.ac.jp
Tanaka
Chikako
C
Yang
Mingying
M
Yao
Juming
J
Kurokawa
Masato
M
D000818:Animals; D001672:Biocompatible Materials / Q000378:metabolism*; D001704:Biopolymers; D012831:Bombyx / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D002455:Cell Division / Q000502:physiology*; D002522:Chlorocebus aethiops; D004548:Elasticity; D005349:Fibroins / Q000235:genetics / Q000378:metabolism; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D008422:Materials Testing / Q000379:methods*; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D011108:Polymers / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D047011:Silk; D014709:Vero Cells
D001672:Biocompatible Materials; D001704:Biopolymers; D005353:Fibronectins; D010455:Peptides; D011108:Polymers; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D047011:Silk; C089172:silk-like protein polymer; C019529:polyalanine; D005349:Fibroins