Isolation and characterization of mannose 6-phosphate/insulin-like growth factor II receptor from bovine serum.
A convenient means was devised for the purification of milligram quantities of a soluble form of the mannose 6-phosphate/insulin-like growth factor II receptor (Man-6-P/IGF II receptor). The receptor was purified to near homogeneity from bovine serum by affinity chromatography on agarose-pentamannosephosphate in the absence of detergent. Approximately 2.5 mg of receptor were obtained from 500 ml of fetal calf serum. The concentration of receptor in serum decreased sharply with development. Fetal calf serum Man-6-P/IGF II receptor was immunologically similar to detergent-solubilized, membrane-bound Man-6-P/IGF II receptor from bovine liver. N-Terminal sequence analysis revealed that the purified serum receptor, but not the solubilized, membrane-associated receptor, contains stoichiometric amounts of bound IGF II. The results of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel chromatography studies suggest that the fetal calf serum receptor (in contrast to the solubilized, membrane-bound bovine testis receptor) does not aggregate. The affinity of the fetal calf serum receptor for bovine testis beta-galactosidase approximated one-half that observed for solubilized, membrane-bound bovine testis receptor.
Li
M
M
Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor 48109.
Distler
J J
JJ
Jourdian
G W
GW
D000375:Aging; D000818:Animals; D002352:Carrier Proteins / Q000097:blood* / Q000302:isolation & purification*; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005333:Fetus; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface / Q000302:isolation & purification* / Q000378:metabolism*; D013737:Testis / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D002352:Carrier Proteins; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface; D001616:beta-Galactosidase