Association of rasGAPSH3 binding protein 1, G3BP1, and rasGap120 with integrin containing complexes induced by an adhesion blocking antibody.
The adhesion blocking antibody 3S3 was used to probe the regulation of alpha5beta1 integrin mediated adhesion in K562 cells. This antibody prevented cellular adherence but it did not interfere with ligand binding by cells or purified integrin. Interaction with 3S3 induced change in the cytoskeletal organization resulting in extensive filopodia formation. The antibody also prevented ligand and anti-integrin antibody induced phosphorylation of FAK in a trans acting fashion. MS based analysis of 3S3 induced integrin containing complexes identified rasGAP SH3 binding protein 1, G3BP1, as a component of these structures. The G3BP1 binding molecule, rasGap120, was also identified in the complexes. Microscopic examination confirmed the recruitment of a component of cellular G3BP1 and rasGap120 pools to sites of integrin cross-linking. G3BP1 was also observed in the 3S3 induced filopodia. In untreated cells, G3BP1 was shown to associate with submembranous regions involved in cellular polarization. Collectively, these results suggest that G3BP1 and rasGap120 can be recruited to sites of integrin ligation where they may play a role in cytoskeletal reorganization. Such changes may result in reduced adhesive potential and account for the 3S3 effects on cellular adhesion. It should be emphasized that these results do not necessarily indicate a direct interaction of integrin with G3BP1 and rasGap120.
Meng
Xiaobo
X
Manitoba Centre for Proteomics, Rheumatic Diseases Research Laboratory and Department of Medicine, University of Manitoba, Winnipeg, Canada.
Krishnan
Jaykumar
J
She
Yemen
Y
Ens
Werner
W
Standing
Kenneth
K
Wilkins
John A
JA
D000906:Antibodies / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D016764:Cell Polarity / Q000187:drug effects / Q000502:physiology; D003599:Cytoskeleton / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism*; D020014:K562 Cells; D013058:Mass Spectrometry; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011554:Pseudopodia / Q000378:metabolism; D020703:ras GTPase-Activating Proteins / Q000378:metabolism*
D000906:Antibodies; D002352:Carrier Proteins; D039081:Integrin alpha5beta1; D020703:ras GTPase-Activating Proteins; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human