Biochemical and structural properties of a Hodgkin's disease-related membrane protein.
The HeFi-1 mAb recognizes a membrane protein on Hodgkin's disease cells and on a limited number of other human cells that are either tumorigenically transformed or virally activated. Herein biochemical and structural analyses of the HeFi-1 reactive membrane protein (HRMP) were done to identify its potential importance in cellular transformation in the Hodgkin's disease cell line L428, in the T cell lymphoma line HuT 78, and in several EBV-transformed lymphoblastoid cell lines. Immunoprecipitation studies demonstrated that the mature form of the HRMP had an apparent Mr of 120 kDa in tumor cells and 116 kDa in the EBV-transformed cell lines and that it was phosphorylated at both serine and tyrosine residues in all cell lines tested. The precursor to the HRMP is an 86-kDa core protein that, after processing by high mannose N-linked glycosylation, migrates with an apparent Mr of 90 kDa. This protein is then further processed to the mature 120-kDa HRMP in part by O-linked glycosylation, the addition of sialic acid residues, and by the conversion of N-linked oligosaccharides from the high mannose to the complex type. Detectable amounts of the 90-kDa molecule can be found in the membrane and, although this protein can be phosphorylated in vitro, it is not phosphorylated in intact cells. The combined results of this study suggest that the HRMP is involved in cellular metabolism and show that an unusual amount of post-translational processing of the 90-kDa precursor results in the formation, and perhaps phosphorylation, of the mature 120-kDa HRMP.
Nawrocki
J F
JF
Department of Medicine, Loyola University Medical Center, Maywood, IL 60153.
Kirsten
E S
ES
Fisher
R I
RI
D000596:Amino Acids / Q000378:metabolism; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D002460:Cell Line; D006689:Hodgkin Disease / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D010750:Phosphoproteins