Post-translational modifications of alpha5beta1 integrin by glycosaminoglycan chains. The alpha5beta1 integrin is a facultative proteoglycan.
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 alpha5beta1 integrin. Using [125I]iodine cell surface labeling or metabolic radiolabeling with sodium [35S]sulfate, we identified alpha5beta1 integrin as the only sulfated integrin among beta1 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 alpha5beta1 integrin sulfation was evidenced by its resistance to treatments with high ionic, chaotrophic, and denaturing agents such as 4 M NaCl, 4 M MgCl2, 8 M urea, and 6 M 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 alpha5beta1 heterodimer and alpha5 and beta1 integrin subunits were proteoglycans. The importance of alpha5beta1 sulfation was strengthened by the finding that this molecule is also sulfated in MG-63 (human osteosarcoma) and HCT-8 (human colon adenocarcinoma) cells.
Veiga
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Ludwig Institute for Cancer Research, R. Prof. Antonio Prudente, 109, 4 A, 01509-010, São Paulo, SP, Brazil.
Elias MCQB
Gremski
W
W
Porcionatto
M A
MA
da Silva
R
R
Nader
H B
HB
Brentani
R R
RR
D002448:Cell Adhesion; D015180:Electrophoresis, Gel, Two-Dimensional; D006025:Glycosaminoglycans / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D017469:Receptors, Fibronectin / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism; D014407:Tumor Cells, Cultured
D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D017469:Receptors, Fibronectin; D013431:Sulfates; C012036:sodium sulfate