Identification of the membrane remnants of transferrin receptor with domain-specific antibodies.
Tissue culture studies with K562 and HL60 cells have demonstrated the production of a soluble form of transferrin receptor identical to that identified in human serum. The present study was undertaken to search for membrane remnants of the truncated receptor with peptide antibodies specific for the extracellular and cytoplasmic domain of transferrin receptor. In cell membranes, a 105K remnant was identified that is consistent with truncation of one extracellular domain monomer of the transferrin receptor. In the exosomal fraction of the culture supernatant, a smaller 20K remnant consistent with truncation of both extracellular domains was also demonstrated. These findings provide evidence that soluble receptor is the product of proteolytic cleavage of intact membrane-bound transferrin receptor. Prior studies showing that the concentration of the extracellular domain in exosomes remained stable during incubation in culture supernatant suggest that this cleavage possibly occurs intracellularly.
Baynes
R D
RD
Department of Medicine and Biochemistry, Kansas University Medical Center, Kansas City 66160-7402.
Shih
Y J
YJ
Hudson
B G
BG
Cook
J D
JD
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000276:immunology; D005110:Extracellular Space / Q000276:immunology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D010449:Peptide Mapping; D011990:Receptors, Transferrin / Q000737:chemistry / Q000276:immunology*
D000906:Antibodies; D010446:Peptide Fragments; D011990:Receptors, Transferrin