Faculty Collaboration Database

Localization of the carbohydrate recognition sites of the insulin-like growth factor II/mannose 6-phosphate receptor to domains 3 and 9 of the extracytoplasmic region. J Biol Chem 2002 Dec 06;277(49):47205-12

Date

10/11/2002

Pubmed ID

12374794

DOI

10.1074/jbc.M208534200

Scopus ID

2-s2.0-0037033019 (requires institutional sign-in at Scopus site)   46 Citations

Abstract

The insulin-like growth factor II/mannose 6-phosphate receptor is a multifunctional receptor that binds to a diverse array of mannose 6-phosphate (Man-6-P) modified proteins as well as nonglycosylated ligands. Previous studies have mapped its two Man-6-P binding sites to a minimum of three domains, 1-3 and 7-9, within its 15-domain extracytoplasmic region. Since the primary amino acid determinants of carbohydrate recognition by the insulin-like growth factor II/mannose 6-phosphate receptor are predicted by sequence alignment to the cation-dependent mannose 6-phosphate receptor to reside within domains 3 and 9, constructs encoding either domain 3 alone or domain 9 alone were expressed in a Pichia pastoris expression system and tested for their ability to bind several carbohydrate ligands, including Man-6-P, pentamannosyl phosphate, the lysosomal enzyme, beta-glucuronidase, and the carbohydrate modifications (mannose 6-sulfate and Man-6-P methyl ester) found on Dictyostelium discoideum lysosomal enzymes. Although both constructs were functional in ligand binding and dissociation, these studies demonstrate the ability of domain 9 alone to fold into a high affinity (K(d) = 0.3 +/- 0.1 nm) carbohydrate-recognition domain whereas the domain 3 alone construct is capable of only low affinity binding (K(d) approximately 500 nm) toward beta-glucuronidase, suggesting that residues in adjacent domains (domains 1 and/or 2) are important, either directly or indirectly, for optimal binding by domain 3.

Author List

Hancock MK, Yammani RD, Dahms NM

Author

Nancy M. Dahms PhD Professor in the Biochemistry department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Binding Sites
Carbohydrate Metabolism
Carbohydrates
Chromatography, Affinity
Cytoplasm
DNA, Complementary
Dictyostelium
Dose-Response Relationship, Drug
Glycosylation
Hexosaminidases
Hydrogen-Ion Concentration
Insulin-Like Growth Factor II
Kinetics
Ligands
Lysosomes
Pichia
Protein Binding
Protein Structure, Tertiary
Receptor, IGF Type 2