The estrogen-inducible transferrin receptor-like membrane glycoprotein is related to stress-regulated proteins.
It was previously shown that estrogen induces a membrane glycoprotein (molecular mass, 95 kDa) in the chicken oviducts, which exhibits several properties similar to transferrin receptors (Poola, I., and Lucas, J. J. (1988) J. Biol. Chem. 263, 19137-19146). In the present study, we have further investigated its molecular and transferrin binding properties. We have sequenced several internal peptides isolated from the purified protein by endopeptidase Lys-C. We have found that it has a high degree of sequence homologies with those of chicken heat-shock protein (cHsp108), mouse endoplasmic reticulum protein (mERp99), hamster glucose-regulated protein (hagrp94), and human tumor rejection antigen (hTRAgp96), all of which are shown to be highly homologous to each other and to yeast hsp90. We demonstrate here that the [35S]methionine-labeled immunoaffinity-purified estrogen-inducible membrane glycoprotein binds to the transferrin affinity columns similar to iron-modulated transferrin receptors. Indirect immunofluorescence microscopic studies indicate that it is an intracellular glycoprotein unlike transferrin receptors. We have isolated two molecular forms of the protein, with molecular masses of 116 and 104 kDa, by immunoaffinity column purification, immunoprecipitation, Western blotting, and pulse-chase labeling analyses. Both 116-and 104-kDa species bind transferrin. This protein can be induced by heat-shocking the oviduct cells at 45 degrees C for 3h and recovering at 37 degrees C for 2-3 h. It is also expressed in the human breast cancer cell lines, MCF-7 and T-47D. All these properties taken together strongly suggest that the estrogen-inducible membrane glycoprotein is a novel transferrin-binding protein, structurally related to the stress-regulated proteins.
Poola
I
I
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Kiang
J G
JG
D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins; D001943:Breast Neoplasms / Q000378:metabolism; D002478:Cells, Cultured; D002645:Chickens; D002846:Chromatography, Affinity; D004967:Estrogens / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006023:Glycoproteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010057:Oviducts / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D011990:Receptors, Transferrin / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014168:Transferrin / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D030161:Avian Proteins; D004967:Estrogens; D006023:Glycoproteins; D006360:Heat-Shock Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011990:Receptors, Transferrin; D014168:Transferrin; C088981:estrogen-inducible transferrin receptor-like membrane glycoprotein, chicken