Design and activity of multifunctional fibrils using receptor-specific small peptides.
We have designed multifunctional peptide fibrils using bioactive laminin-derived peptides and evaluated their potential as a biomedical material for tissue engineering. The Leu-Arg-Gly-Asp-Asn (LRGDN) peptide derived from laminin-111, which contains an RGD sequence bound to integrin alphavbeta3, was added to the N-terminus of the four amyloidogenic cell-adhesive laminin-derived peptides (A119: LSNIDYILIKAS, AG97: SAKVDAIGLEIV, B133: DISTKYFQMSLE, and B160: VILQQSAADIAR). The RGD-conjugated peptides were stained with Congo red and exhibited amyloid-like fibril formation in the electron microscopic. The RGD-conjugated peptides promoted human dermal fibroblasts spreading with well-organized actin stress fibers and focal contacts. Human dermal fibroblast attachment to the RGD-conjugated peptides was inhibited by anti-alphav integrin antibody. Further, cell attachment to B133 was inhibited by anti-alpha2 and anti-beta1 integrin antibodies, whereas attachment to RGD-B133 was inhibited by anti-alphav and anti-beta1 integrin antibodies. These results suggest that the RGD-conjugated peptides interact with integrin alphavbeta3 and that RGD-B133 interacts with both integrin alphavbeta3 and integrin beta1. The RGD-conjugated peptide fibrils promoted neurite outgrowth in a peptide-dependent manner. These results support that biologically active sequence-conjugated peptide fibrils interact in a receptor-specific manner with cells and promote multifunctional activities. These fibrils may have use as biological supports for cell-specific tissue engineering.
Ohga
Yukiko
Y
Laboratory of Clinical Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan.
Katagiri
Fumihiko
F
Takeyama
Kazuki
K
Hozumi
Kentaro
K
Kikkawa
Yamato
Y
Nishi
Norio
N
Nomizu
Motoyoshi
M
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000682:Amyloid / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004396:Coloring Agents / Q000378:metabolism; D003224:Congo Red / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007231:Infant, Newborn; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D019012:Integrin beta1 / Q000378:metabolism; D007797:Laminin / Q000737:chemistry; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D016716:PC12 Cells; D051381:Rats; D012867:Skin / Q000166:cytology
D000199:Actins; D000682:Amyloid; D004396:Coloring Agents; D039302:Integrin alphaVbeta3; D019012:Integrin beta1; D007797:Laminin; D009842:Oligopeptides; C109085:RGDN peptide; D003224:Congo Red; C047981:arginyl-glycyl-aspartic acid