Characterization of heparin-binding growth-associated factor receptor on NIH 3T3 cells.
Scatchard plot analysis of the binding of 125I-labeled heparin binding cell growth-associated factor (125I-HBGAF) to NIH 3T3 cells revealed a single class of high affinity receptors (-5000/cell) with kd of -0.6 nM. 125I-HBGAF was covalently cross-linked to the cell surface receptor on NIH 3T3 cells with disuccinimidyl suberate (DSS). Two 125I-HBGAF-cross-linked complexes of 170 kDa and 142 kDa were observed on SDS-polyacrylamide gel electrophoresis under reducing and nonreducing conditions. The 125I-HBGAF-cross-linked complex formation was completely abolished in the presence of greater than or equal to 100-fold excess of unlabeled HBGAF but not PDGF, EGF, aFGF, bFGF, or insulin. 125I-HBGAF appeared to undergo rapid internalization and relatively slow degradation following binding to the HBGAF receptor on NIH 3T3 cells. These results suggest that NIH 3T3 cells express a high affinity HBGAF receptor which shows two different estimated molecular masses of -155 kDa and -127 kDa. This high affinity HBGAF receptor was also found to express in other cell types.
Kuo
M D
MD
Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, MO 63104.
Huang
S S
SS
Huang
J S
JS
D016475:3T3 Cells; D000818:Animals; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism*; D016228:Endothelial Growth Factors / Q000378:metabolism; D006133:Growth Substances / Q000378:metabolism*; D007457:Iodine Radioisotopes; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D011869:Radioligand Assay; D011975:Receptors, Mitogen / Q000302:isolation & purification / Q000378:metabolism*; D040262:Receptors, Vascular Endothelial Growth Factor
D002352:Carrier Proteins; D016207:Cytokines; D016228:Endothelial Growth Factors; D006133:Growth Substances; D007457:Iodine Radioisotopes; D011975:Receptors, Mitogen; C066845:pleiotrophin; D040262:Receptors, Vascular Endothelial Growth Factor