Glycosaminoglycan chains from alpha5beta1 integrin are involved in fibronectin-dependent cell migration.
Alpha5beta1 integrin from both wild-type CHO cells (CHO-K1) and deficient in proteoglycan biosynthesis (CHO-745) is post-translationally modified by glycosaminoglycan chains. We demonstrated this using [35S]sulfate metabolic labeling of the cells, enzymatic degradation, immunoprecipitation reaction with monoclonal antibody, fluorescence microscopy, and flow cytometry. The alpha5beta1 integrin heterodimer is a hybrid proteoglycan containing both chondroitin and heparan sulfate chains. Xyloside inhibition of sulfate incorporation into alpha5beta1 integrin also supports that integrin is a proteoglycan. Also, cells grown with xyloside adhered on fibronectin with no alteration in alpha5beta1 integrin expression. However, haptotactic motility on fibronectin declined in cells grown with xyloside or chlorate as compared with controls. Thus, alpha5beta1 integrin is a proteoglycan and the glycosaminoglycan chains of the integrin influence cell motility on fibronectin. Similar glycosylation of alpha5beta1 integrin was observed in other normal and malignant cells, suggesting that this modification is conserved and important in the function of this integrin. Therefore, these glycosaminoglycan chains of alpha5beta1 integrin are involved in cellular migration on fibronectin.
Franco
Célia R C
CR
Departamento de Bioquímica, Universidade Federal de São Paulo, Rua Três de Maio, 100-CEP 04044-020, São Paulo, SP, Brazil.
Trindade
Edvaldo S
ES
Rocha
Hugo A O
HA
da Silveira
Rafael Bertoni
RB
Paludo
Katia Sabrina
KS
Chammas
Roger
R
Veiga
Silvio S
SS
Nader
Helena B
HB
Dietrich
Carl P
CP
D000818:Animals; D016466:CHO Cells; D002465:Cell Movement / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D004587:Electrophoresis, Agar Gel; D005353:Fibronectins / Q000502:physiology*; D005434:Flow Cytometry; D006025:Glycosaminoglycans / Q000737:chemistry*; D047468:Immunoprecipitation; D039081:Integrin alpha5beta1 / Q000737:chemistry*; D008856:Microscopy, Fluorescence
D005353:Fibronectins; D006025:Glycosaminoglycans; D039081:Integrin alpha5beta1