Biochemical evidence that the type II insulin-like growth factor receptor is identical to the cation-independent mannose 6-phosphate receptor.
Cloning and sequencing of the human type II insulin-like growth factor (IGF) receptor cDNA revealed an 80% deduced amino acid sequence homology with the bovine cation-independent mannose 6-phosphate (Man-6-P) receptor, suggesting identity of the two receptors (Morgan, D. O., Edman, J. C., Standring, D. N., Fried, V. A., Smith, M. C., Roth, R. A., and Rutter, W. J. (1987) Nature 329, 301-307). We have performed biochemical experiments that support this proposal. Rat liver type II IGF receptor, purified by the conventional method of IGF-II affinity chromatography, bound quantitatively to a beta-galactosidase affinity column and was eluted with Man-6-P. Bovine liver Man-6-P receptor, prepared by the conventional method of affinity chromatography on phosphomannan-Sepharose, bound IGF-II with high affinity (Kd = 1 nM). Affinity cross-linking of 125I-IGF-II to the Man-6-P receptor and analysis by sodium dodecyl sulfate-gel electrophoresis showed that beta-galactosidase, but not Man-6-P, inhibited the formation of the 250-kDa 125I-IGF-II-receptor complex. The inhibition by beta-galactosidase was prevented by coincubation with Man-6-P. 125I-IGF-II did not bind to the 46-kDa cation-dependent Man-6-P receptor. For immunologic studies we purified type II IGF receptors and Man-6-P receptors in parallel from rat placental membranes using either IGF-II- or beta-galactosidase affinity chromatography. A panel of five antisera that previously had been raised against either type II IGF receptor or Man-6-P receptor behaved identically toward type II IGF receptor versus Man-6-P receptor in ligand blocking and immunoprecipitation assays. Our data support the conclusion that the type II IGF receptor and the cation-independent Man-6-P receptor are the same protein and that the IGF-II and Man-6-P-binding sites are distinct.
Kiess
W
W
Endocrinology Section, National Cancer Institute, Bethesda, Maryland 20892.
Blickenstaff
G D
GD
Sklar
M M
MM
Thomas
C L
CL
Nissley
S P
SP
Sahagian
G G
GG
D000818:Animals; D002352:Carrier Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002846:Chromatography, Affinity; D005260:Female; D006600:Hexosephosphates / Q000378:metabolism*; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008360:Mannosephosphates / Q000378:metabolism*; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017527:Receptor, IGF Type 2; D011972:Receptor, Insulin / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D017451:Receptors, Somatomedin; D013002:Somatomedins / Q000378:metabolism*; D013329:Structure-Activity Relationship
D002352:Carrier Proteins; D006600:Hexosephosphates; D008360:Mannosephosphates; D017527:Receptor, IGF Type 2; D017451:Receptors, Somatomedin; D013002:Somatomedins; D007335:Insulin-Like Growth Factor II; D011972:Receptor, Insulin