Alpha 4/180, a novel form of the integrin alpha 4 subunit.
The integrin alpha 4 beta 1 (VLA-4) is a versatile cell-cell and cell-extracellular matrix adhesion receptor. The alpha 4 subunit can be expressed on the cell surface in two forms: an intact form (alpha 4/150) and a cleaved form (alpha 4/80,70). Here we have characterized a third form of alpha 4, called alpha 4/180. Although alpha 4/180 (M(r) 180, nonreduced) is different in size than alpha 4/150 (M(r) 150, nonreduced), these two forms are clearly related, as they (i) shared the same amino-terminal sequence, (ii) were both recognized in Western blots by an anti-alpha 4 COOH-terminal antiserum, (iii) migrated with the same apparent size and charge when reduced, and (iv) were both immunoprecipitated using anti-VLA-4 reagents. In pulse-chase experiments, precursors to both forms appeared simultaneously and matured at the same rate, indicating that one is most likely not the biosynthetic precursor of the other. Although reduction of alpha 4/180 to yield alpha 4/150 suggested the release of a cysteine-linked 30-kDa fragment, seven different biochemical techniques failed to identify such a fragment. Also, alpha 4/180 was converted to alpha 4/150 by incubation at pH 11, by treatment with EDTA at 56 degrees C, or by heating in the presence of elevated SDS levels. Together our findings suggest that alpha 4/180 and alpha 4/150 represent different conformations of the same alpha 4 polypeptide, with the former having anomalous slower migration in SDS-polyacrylamide gel electrophoresis. This unusual biochemical feature of alpha 4 is not shared by other beta 1-associated integrin alpha subunits and suggests that VLA-4 has unique structural properties.
Parker
C M
CM
Brigham and Womens Hospital, Department of Rheumatology and Immunology, Boston, Massachusetts.
Pujades
C
C
Brenner
M B
MB
Hemler
M E
ME
D017398:Alternative Splicing; D015153:Blotting, Western; D002460:Cell Line; D004220:Disulfides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D039001:Integrin alpha Chains; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D016029:Receptors, Very Late Antigen / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
C063493:CDw49d; D004220:Disulfides; D039001:Integrin alpha Chains; D016029:Receptors, Very Late Antigen