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J Biol Chem, Vol. 274, Issue 38, 27076-27082, September 17, 1999

The Kangaroo Cation-independent Mannose 6-Phosphate Receptor Binds Insulin-like Growth Factor II with Low Affinity

Catherine A. YandellDagger §, Andrew J. DunbarDagger §, John F. WheldrakeDagger , and Zee UptonDagger §

From the Dagger  Cooperative Research Centre for Tissue Growth and Repair, P. O. Box 10065, Adelaide B.C., South Australia, Australia, 5000, § CSIRO Human Nutrition, P. O. Box 10041, Adelaide B.C., South Australia, Australia, 5000, and  School of Biological Sciences, Flinders University of South Australia, GPO 2100, Adelaide, South Australia, Australia, 5001

The mammalian cation-independent mannose 6-phosphate receptor (CI-MPR) binds mannose 6-phosphate-bearing glycoproteins and insulin-like growth factor (IGF)-II. However, the CI-MPR from the opossum has been reported to bind bovine IGF-II with low affinity (Dahms, N. M., Brzycki-Wessell, M. A., Ramanujam, K. S., and Seetharam, B. (1993) Endocrinology 133, 440-446). This may reflect the use of a heterologous ligand, or it may represent the intrinsic binding affinity of this receptor. To examine the binding of IGF-II to a marsupial CI-MPR in a homologous system, we have previously purified kangaroo IGF-II (Yandell, C. A., Francis, G. L., Wheldrake, J. F., and Upton, Z. (1998) J. Endocrinol. 156, 195-204), and we now report the purification and characterization of the CI-MPR from kangaroo liver. The interaction of the kangaroo CI-MPR with IGF-II has been examined by ligand blotting, radioreceptor assay, and real-time biomolecular interaction analysis. Using both a heterologous and homologous approach, we have demonstrated that the kangaroo CI-MPR has a lower binding affinity for IGF-II than its eutherian (placental mammal) counterparts. Furthermore, real-time biomolecular interaction analysis revealed that the kangaroo CI-MPR has a higher affinity for kangaroo IGF-II than for human IGF-II. The cDNA sequence of the kangaroo CI-MPR indicates that there is considerable divergence in the area corresponding to the IGF-II binding site of the eutherian receptor. Thus, the acquisition of a high-affinity binding site for regulating IGF-II appears to be a recent event specific to the eutherian lineage.


Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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