Silklike materials constructed from sequences of Bombyx mori silk fibroin, fibronectin, and elastin.
Two silklike proteins, [TGRGDSPAGG(GAGAGS)3AS]5 (FS5) and [TGRGDSPA-(GVPGV)2GG(GAGAGS)3AS]8 (FES8) were designed to demonstrate the superior performance as biomaterials of silklike proteins. The former protein consists of the crystalline domain sequence, (GAGAGS)n from Bombyx mori silk fibroin and cell-adhesive sequence TGRGDSPA coming from fibronectin-containing RGD triplet. The additional sequence (GVPGV)n from elastin was included in the latter protein. The considerably higher cell-adhesion activities of these proteins for NHDF and VERO cells were observed by comparing with those of silklike materials without RGD sequences and also the crystalline fraction of B. mori silk fibroin. This tendency was independent of the treatments, 4.5M LiClO4 or formic acid (FA), on silklike proteins. Their activities are also higher than those of commercial Fibronectin F for NHDF cell. Their structural characterization was studied using 13C solid-state NMR. Although the overlapped peaks in usual 13C CP/MAS NMR spectra make the detailed structural analysis difficult, the methyl resonance regions observed using dipolar dephasing NMR were very useful for the analysis. The presence of both random coil and beta-sheet structures was observed in these proteins clearly. The content of beta-sheet structure in both proteins increases after FA treatment when compared with the lyophilized samples. The production of electrospun nanofibers from their hexafluoroacetone solution was also tried. The silklike protein FES8 could prepare nonwoven silk fibers although FS5 could not.
Yang
Mingying
M
Department of Biotechnology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan.
Tanaka
Chikako
C
Yamauchi
Kazuo
K
Ohgo
Kosuke
K
Kurokawa
Masato
M
Asakura
Tetsuo
T
D000096:Acetone / Q000031:analogs & derivatives / Q000737:chemistry; D000818:Animals; D001483:Base Sequence; D001672:Biocompatible Materials / Q000737:chemistry; D012831:Bombyx / Q000737:chemistry*; D002448:Cell Adhesion / Q000187:drug effects; D002522:Chlorocebus aethiops; D004549:Elastin / Q000737:chemistry* / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism; D005349:Fibroins / Q000737:chemistry* / Q000235:genetics*; D005353:Fibronectins / Q000737:chemistry* / Q000235:genetics*; D005466:Fluorocarbons / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D043942:Nanotubes; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D047011:Silk / Q000737:chemistry* / Q000235:genetics*; D054457:Tissue Scaffolds; D014709:Vero Cells; D006845:Hydrocarbons, Fluorinated
D000096:Acetone; D001672:Biocompatible Materials; D004549:Elastin; D005349:Fibroins; D005353:Fibronectins; D005466:Fluorocarbons; D011994:Recombinant Proteins; D047011:Silk; C017366:hexafluoroacetone; D006845:Hydrocarbons, Fluorinated