Purification of a new type high molecular weight receptor (type V receptor) of transforming growth factor beta (TGF-beta) from bovine liver. Identification of the type V TGF-beta receptor in cultured cells.
A 400-kDa transforming growth factor beta (TGF-beta) receptor was purified from plasma membranes of bovine liver using Triton X-100 extraction, wheat germ lectin-Sepharose 4B affinity chromatography, DEAE-cellulose anion exchange chromatography, and Sepharose CL-4B gel filtration chromatography. This procedure yielded approximately 20 micrograms of the receptor from 1 kg of bovine liver. During purification, the 400-kDa TGF-beta receptor was detected by a cross-linking assay in which the TGF-beta receptor-125I-TGF-beta complex was cross-linked by disuccinimidyl suberate, a bifunctional reagent, and analyzed by 5.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography. This novel 400-kDa TGF-beta receptor was also identified on cultured cells including cells reported to lack the type III receptor. The 400-kDa TGF-beta receptor, a nonproteoglycan glycoprotein, appears to be distinct from TGF-beta receptors (types I, II, III, and IV) previously identified on cultured cells and is designated as the type V receptor. The 400-kDa TGF-beta receptor as well as type I, II, and III receptors underwent internalization upon 125I-TGF-beta binding in mink lung epithelial cells.
O'Grady
P
P
Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, Missouri 63104.
Kuo
M D
MD
Baldassare
J J
JJ
Huang
S S
SS
Huang
J S
JS
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D004847:Epithelial Cells; D008099:Liver / Q000737:chemistry* / Q000378:metabolism; D008907:Mink; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011956:Receptors, Cell Surface / Q000302:isolation & purification* / Q000378:metabolism; D018125:Receptors, Transforming Growth Factor beta; D013388:Succinimides / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000378:metabolism
D003432:Cross-Linking Reagents; D011956:Receptors, Cell Surface; D018125:Receptors, Transforming Growth Factor beta; D013388:Succinimides; D016212:Transforming Growth Factor beta; C019358:disuccinimidyl suberate