Improving cell-adhesive properties of recombinant Bombyx mori silk by incorporation of collagen or fibronectin derived peptides produced by transgenic silkworms.
Silk of Bombyx mori can be used as various biomaterials. Especially, it is useful as a protein for coating the surface of cell culture plates since the silk possesses a biocompatibility to the cultured cells. However, the cell-adhesive ability is weaker than collagen or fibronectin, which are used for coating the plate more frequently (Yao et al. J. Biochem., 2004, 136, 643-649). To increase the biocompatibility of the silk, we constructed transgenic silkworms, inserting the modified fibroin light-chain genes for making recombinant silks that possessed partial collagen or fibronectin sequences, that is, [GERGDLGPQGIAGQRGVV(GER)3GAS]8GPPGPCCGGG or [TGRGDSPAS]8, respectively. Films were made from the recombinant silks, and the cell-adhesive activity for cultured mammalian cells was observed. The results showed that the two types of recombinant silk films possessed a much higher cell-adhesive activity as compared to the original unmodified silk. Especially, the recombinant silk with the sequence [TGRGDSPAS]8, produced by a transgenic Nd-sD mutant, gave a 6 times higher activity than the original unmodified silk.
Yanagisawa
Satoshi
S
Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan.
Zhu
Zhenghua
Z
Kobayashi
Isao
I
Uchino
Keiro
K
Tamada
Yasushi
Y
Tamura
Toshiki
T
Asakura
Tetsuo
T
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D003094:Collagen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005353:Fibronectins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D047011:Silk / Q000235:genetics / Q000378:metabolism*
D005353:Fibronectins; D009842:Oligopeptides; D010446:Peptide Fragments; D011994:Recombinant Proteins; D047011:Silk; C047981:arginyl-glycyl-aspartic acid; D003094:Collagen