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Title: Post-translational modifications of alpha(5)beta(1) integrin by glycosaminoglycan chains - the alpha(5)beta(1) integrin is a facultative proteoglycan
Authors: Veiga, Silvio Sanches [UNIFESP]
Elias, MCQB
Gremski, W.
Porcionatto, Marimélia Aparecida [UNIFESP]
Silva, Roseli da [UNIFESP]
Nader, Helena Bonciani [UNIFESP]
Brentani, Ricardo Renzo [UNIFESP]
Universidade Federal de São Paulo (UNIFESP)
Issue Date: 9-May-1997
Publisher: Amer Soc Biochemistry Molecular Biology Inc
Citation: Journal of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 272, n. 19, p. 12529-12535, 1997.
Abstract: Cell-fibronectin interactions, mediated through several different receptors, have been implicated in a wide variety of cellular properties. Among the cell surface receptors for fibronectin, integrins are the best characterized, particularly the prototype alpha(5) beta(1) integrin. Using [I-125]iodine cell surface labeling or metabolic radiolabeling with sodium [S-35]sulfate, we identified alpha(5) beta(1) integrin as the only sulfated integrin among beta(1) integrin heterodimers expressed by the human melanoma cell line Mel-85. This facultative sulfation was confirmed not only by immunoprecipitation reactions using specific monoclonal antibodies but also by fibronectin affinity chromatography, two dimensional electrophoresis, and chemical reduction, the covalent nature of alpha(5) beta(1) integrin sulfation was evidenced by its resistance to treatments with high ionic, chaotrophic, and denaturing agents such as 4 nz NaCl, 4 hn MgCl2, 8 M urea, and 6 ar guanidine HCl. Based on deglycosylation procedures as chemical beta-elimination, proteinase K digestion, and susceptibility to glycosaminoglycan lyases (chondroitinase ABC and heparitinases I and II), it was demonstrated that the alpha(5) beta(1) heterodimer and alpha(5) and beta(1) integrin subunits were proteoglycans. the importance of alpha(5) beta(1) sulfation was strengthened by the finding that this molecule is also sulfated in MG-63 (human osteosarcoma) and HCT-8 (human colon adenocarcinoma) cells.
ISSN: 0021-9258
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