Membrane proteins from lymphoblastoid cells showing cross-affinity for alpha-fetoprotein and albumin. Isolation and characterization.
AFP or SA immobilized on nitrocellulose membranes (AFP-NC or SA-NC) were used as affinity matrices to purify cell membrane proteins with affinity for AFP (AFP-BP) and for SA (SA-BP) from membrane-enriched extracts of Raji cells (a B-lymphoma cell line), as well as for normal resting and activated peripheral blood lymphocytes (PBMC). SDS-PAGE and ligand blotting assays showed that AFP-BP and SA-BP isolated from Raji cells are probably identical molecules. They consisted of two sets of polypeptides of 31 kDa and 18 kDa. The glycoprotein nature of isolated 31 kDa and 18 kDa peptides was suggested by positive staining with Schiff's reagent, and amino-acid analysis revealed similar amino-acid composition for the two glycoproteins. In human PHA-activated PBMC, only the 18 kDa polypeptide was identified and isolated as AFP-BP or SA-BP. As in Raji cells, this 18 kDa polypeptide, isolated by affinity for AFP or for SA, appeared to be the same molecule. Contrary to Raji cells and activated PBMC, no proteins with an affinity for AFP or for SA were identified or isolated in resting PBMC. These observations strongly suggest that the isolated 31 kDa and 18 kDa glycoproteins are probably AFP receptors previously demonstrated in several neoplastic and normal cells undergoing growth and/or differentiation; indeed, they were identical to albumin-binding proteins described by others.
Torres
J M
JM
Laboratorie de Chimie des Proteines, Institut de Recherches Scientifiques sur le Cancer (CNRS), Villejuif, France.
Darracq
N
N
Uriel
J
J
D000596:Amino Acids / Q000032:analysis; D002462:Cell Membrane / Q000737:chemistry; D006801:Humans; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000378:metabolism*
D000596:Amino Acids; D008562:Membrane Glycoproteins; D012709:Serum Albumin; D000509:alpha-Fetoproteins