Proteolytic cleavage of the integrin beta 4 subunit.
The integrin beta 4 subunit often undergoes proteolytic cleavage within its long cytoplasmic tail to yield a characteristic protein pattern of 205, 165, and 125 kDa. The results in this study suggest that beta 4 cleavage often occurs during or after cell lysis, where it was readily inhibitable by calcium chelators (EDTA, EGTA) and inhibitors of cysteine proteases (E64c, leupeptin). The cleavage of beta 4 is catalyzed by a calpain-like enzyme because (i) it requires calcium, (ii) it is mimicked by purified milli-calpain, and (iii) it is inhibited by several calpain inhibitors including the calpain-specific inhibitor calpastatin. Within intact cells, cleavage of beta 4 was cell type-specific and observed only when the cells were made permeable to calcium. Substantial cell viability was retained during beta 4 cleavage induced by ionomycin plus calcium, indicating that cleavage within intact cells was not necessarily a consequence of cell death. However, manipulations of cells including suspension, synchronization, and stimulation with serum, phorbol esters, or other agents all failed to induce cleavage, suggesting that if cleavage is physiologically relevant, it is not easily duplicated in vitro. Analysis of multiple cell types showed a wide variation in beta 4 sensitivity to proteolytic cleavage, suggesting that this process might be differentially regulated depending on the cellular environment.
Potts
A J
AJ
Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115.
Croall
D E
DE
Hemler
M E
ME
D000954:Antigens, Surface / Q000302:isolation & purification; D016477:Artifacts; D002118:Calcium / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000494:pharmacology; D002154:Calpain / Q000037:antagonists & inhibitors / Q000494:pharmacology; D002448:Cell Adhesion; D002470:Cell Survival; D004072:Digitonin / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D006801:Humans; D039161:Integrin alpha6beta4; D039663:Integrin beta4; D016023:Integrins / Q000302:isolation & purification / Q000378:metabolism*; D015759:Ionomycin / Q000494:pharmacology; D010446:Peptide Fragments / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; D002135:Calcium-Binding Proteins; D039161:Integrin alpha6beta4; D039663:Integrin beta4; D016023:Integrins; D010446:Peptide Fragments; D004533:Egtazic Acid; D015759:Ionomycin; C033668:calpastatin; D010450:Endopeptidases; D002154:Calpain; D004072:Digitonin; D002118:Calcium