Possible involvement of altered RGD sequence in reduced adhesive and spreading activities of advanced glycation end product-modified fibronectin to vascular smooth muscle cells.
Although fibronectin (FN) modified by advanced glycation end products (AGEs) has been shown to contribute to the development of diabetic vascular complications through its reduced adhesive activity to vascular cells, little is known about changes in the cell binding domain of AGE-modified FN. Here we examined the mechanism of reduced adhesive and spreading activities of AGE-modified FN to vascular smooth muscle cells (SMCs), particularly the contribution of modification of Arg-Gly-Asp (RGD) sequence. Incubation with glucose caused not only the formation of N(epsilon) -carboxymethyllysine and pentosidine, but also polymerization of FN in a dose- and time-dependent manner. AGE-modified FN had significantly low adhesive and spreading activities to cultured SMCs. On the other hand, multimeric FN formed by disulfide bonds did not show any effect on either cell adhesion or spreading. The adhesive activity of type I collagen, one of the RGD sequence-containing proteins, to SMCs also decreased by AGE-modification. The inhibitory effect of AGE-modification on cell adhesion was significantly greater in type I collagen than in FN. Although the extent of AGE-modification of type I collagen was indistinguishable from that of FN, AGE-modification decreased the arginine content of type I collagen by 69.5% and of FN by 30.6%, compared with their non-glycated forms. The addition of RGD peptides caused a decrease in adhesion of SMCs to non-glycated FN, but not to AGE-modified FN. Modification of RGD sequence with glyoxal eliminated its inhibitory effect on cell adhesion. Our results suggest that a marked decrease in adhesive and spreading activities of AGE-modified FN to SMCs might largely be due to a modification of its RGD sequence by AGE, thus suggesting a potential link between AGE modification of FN and the pathogenesis of diabetic angiopathy.
Sakata
N
N
Department of Pathology, School of Medicine; Fukuoka University, 45-1, 7-chome Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan. nysakata@fukuoka-u.ac.jp
Sasatomi
Y
Y
Meng
J
J
Ando
S
S
Uesugi
N
N
Takebayashi
S
S
Nagai
R
R
Horiuchi
S
S
D000596:Amino Acids; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005353:Fibronectins / Q000276:immunology / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D007120:Immunochemistry / Q000379:methods; D008239:Lysine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000276:immunology; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D009842:Oligopeptides / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate; D013552:Swine
D000596:Amino Acids; D005353:Fibronectins; D017127:Glycation End Products, Advanced; D009842:Oligopeptides; D012967:Sodium Dodecyl Sulfate; C048496:N(6)-carboxymethyllysine; C047981:arginyl-glycyl-aspartic acid; D003094:Collagen; D001120:Arginine; C062187:pentosidine; D008239:Lysine