The insulin-like growth factor II/mannose-6-phosphate receptor is present in monkey serum.
We recently reported that the insulin-like growth factor II (IGF-II)/mannose-6-phosphate (Man-6-P) receptor is present in fetal and postnatal rat serum and that its serum content declined dramatically postnatally between days 20 and 40 . We now provide evidence that the IGF-II/Man-6-P receptor is also present in monkey serum. Serum was gel filtered on Sephadex G-200, and the column fractions were assayed for binding of radiolabeled IGF-II. There was significant binding of [125I]IGF-II to the void volume fractions in addition to binding to the 150K and 40K carrier proteins. Binding to the void volume fractions was greatest in cord serum and decreased with age. Competitive binding studies with [125I]IGF-II and the void volume pools from monkey serum demonstrated that IGF-I competed less potently than IGF-II, and insulin did not compete. Radiolabeled IGF-I did not bind specifically to the void volume pools. Chemical cross-linking of [125I]IGF-II to aliquots of the void volume pools from monkey cord serum samples and analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of dithiothreitol demonstrated a specific band at about 240K. Western blotting using a specific antiserum (no. 3637) against rat IGF-II/Man-6-P receptor was performed on aliquots of the Sephadex G-200 void volume pools of monkey serum. A band of approximately the same size as that found with human fibroblast members (approximately 215 K without dithiothreitol) was detected. The IGF-II/Man-6-P receptor band was more intense in cord serum than in the postnatal samples. When cord serum Sephadex G-200 pools were gel filtered on Sephadex G-50 in 1 mol/L acetic acid to separate binding components from free IGF, and IGF-II was measured by RRA, approximately 20% of the circulating IGF-II was found to be associated with this IGF-II/Man-6-P receptor in monkey serum. We conclude that the IGF-II/Man-6-P receptor present in serum may be a significant carrier for IGF-II in the monkey.
Gelato
M C
MC
Metabolism Branch, National Cancer Institute, Bethesda, Maryland 20892.
Kiess
W
W
Lee
L
L
Malozowski
S
S
Rechler
M M
MM
Nissley
P
P
D000375:Aging / Q000097:blood; D000818:Animals; D002352:Carrier Proteins / Q000097:blood*; D002850:Chromatography, Gel; D005312:Fetal Blood / Q000032:analysis*; D005347:Fibroblasts / Q000032:analysis; D006801:Humans; D007158:Immunologic Techniques; D007335:Insulin-Like Growth Factor II / Q000097:blood*; D008252:Macaca fascicularis; D008253:Macaca mulatta; D011869:Radioligand Assay; D017527:Receptor, IGF Type 2; D011972:Receptor, Insulin / Q000032:analysis*; D017451:Receptors, Somatomedin; D013002:Somatomedins / Q000097:blood*
D002352:Carrier Proteins; D017527:Receptor, IGF Type 2; D017451:Receptors, Somatomedin; D013002:Somatomedins; D007335:Insulin-Like Growth Factor II; D011972:Receptor, Insulin