Lymphotoxin is expressed as a heteromeric complex with a distinct 33-kDa glycoprotein on the surface of an activated human T cell hybridoma.
We characterized the membrane-associated form of lymphotoxin (surface LT) on the activated II-23.D7 T cell hybridoma. Antibodies to rLT precipitated both surface LT and a distinct 33-kDa glycoprotein (p33). Because p33 and surface LT were antigenically unrelated, their coprecipitation suggested a physical association of p33 and surface LT on the membrane. Pulse-chase analysis indicated that LT and p33 associate with each other early in the LT biosynthetic pathway, precluding the possibility that LT is secreted and bound to p33 or a surface receptor. Furthermore, no p33 was associated with the secreted form of LT. Isoelectric focusing of surface LT and p33 under nondenaturing and denaturing conditions confirmed that surface LT and p33 existed as a complex. Treatment of cells with a high concentration of salt or with acid indicated that surface LT is a peripheral membrane protein. Although secreted LT is a homologous trimer, protein cross-linking studies revealed that surface LT existed as a monomer associated with a dimer of p33. Together the results demonstrate a novel mechanism for stable membrane expression of LT by activated T cells.
Androlewicz
M J
MJ
Division of Biomedical Sciences, University of California, Riverside 92521-0121.
Browning
J L
JL
Ware
C F
CF
D000940:Antigenic Variation; D001345:Autoradiography; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism*; D007525:Isoelectric Focusing; D008233:Lymphotoxin-alpha / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011233:Precipitin Tests; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*
D003432:Cross-Linking Reagents; D008233:Lymphotoxin-alpha; D008562:Membrane Glycoproteins