Skip to main content

An insulin-like growth factor II (IGF-II) affinity-enhancing domain localized within extracytoplasmic repeat 13 of the IGF-II/mannose 6-phosphate receptor.

Publication ,  Journal Article
Devi, GR; Byrd, JC; Slentz, DH; MacDonald, RG
Published in: Mol Endocrinol
November 1998

Insulin-like growth factor II (IGF-II) and phosphomannosylated glycoproteins bind to distinct sites on the same receptor, the IGF-II/mannose 6-phosphate receptor (IGF2R). Analysis of truncated receptors (minireceptors) has been used to map the IGF-II binding site within the receptor's extracytoplasmic domain, which consists of 15 homologous repeats. A minireceptor consisting of repeat 11 contained the minimal elements for binding IGF-II, but with 5- to 10-fold lower relative binding affinity than the full-length receptor. We hypothesized that the complete, high-affinity IGF-II binding site is formed by interaction between the primary site in repeat 11 and a putative affinity-enhancing domain. To determine the minimum portion of the IGF2R's extracytoplasmic domain needed for expression of high-affinity IGF-II binding, a nested set of FLAG epitope-tagged minireceptors encompassing repeats 11 through 15 was prepared and transiently expressed in 293T cells. Minireceptors containing repeats 11-13 or 11-15 exhibited high affinity, comparable to the full-length receptor (IC50 = 1-2 nM), whereas constructs containing repeat 11 only or repeats 11-12 did not (IC50 = 10-20 nM). These data suggested that the affinity-enhancing domain is located within repeat 13, which contains a unique 43-residue insert that has approximately 50% sequence identity to the type II repeat of fibronectin. Although a repeat 13 minireceptor did not bind IGF-II on its own, an 11-13 minireceptor containing a deletion of the 43-residue insert exhibited low IGF-II binding affinity (IC50 = 10-20 nM). Expression of mutant receptors from a full-length IGF2R construct bearing a deletion of the 43-residue insert was very low relative to wild type. Depletion assays using IGF-II-Sepharose showed that the mutant receptor had lower affinity for IGF-II than the wild-type receptor. This study reveals that two independent receptor domains are involved in the formation of a high-affinity binding site for IGF-II, and that a complete repeat 13 is required for high-affinity IGF-II binding.

Duke Scholars

Published In

Mol Endocrinol

DOI

ISSN

0888-8809

Publication Date

November 1998

Volume

12

Issue

11

Start / End Page

1661 / 1672

Location

United States

Related Subject Headings

  • Structure-Activity Relationship
  • Repetitive Sequences, Amino Acid
  • Recombinant Fusion Proteins
  • Receptor, IGF Type 2
  • Protein Structure, Tertiary
  • Protein Binding
  • Insulin-Like Growth Factor II
  • Humans
  • Genes, Synthetic
  • Endocrinology & Metabolism
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Devi, G. R., Byrd, J. C., Slentz, D. H., & MacDonald, R. G. (1998). An insulin-like growth factor II (IGF-II) affinity-enhancing domain localized within extracytoplasmic repeat 13 of the IGF-II/mannose 6-phosphate receptor. Mol Endocrinol, 12(11), 1661–1672. https://doi.org/10.1210/mend.12.11.0192
Devi, G. R., J. C. Byrd, D. H. Slentz, and R. G. MacDonald. “An insulin-like growth factor II (IGF-II) affinity-enhancing domain localized within extracytoplasmic repeat 13 of the IGF-II/mannose 6-phosphate receptor.Mol Endocrinol 12, no. 11 (November 1998): 1661–72. https://doi.org/10.1210/mend.12.11.0192.
Devi, G. R., et al. “An insulin-like growth factor II (IGF-II) affinity-enhancing domain localized within extracytoplasmic repeat 13 of the IGF-II/mannose 6-phosphate receptor.Mol Endocrinol, vol. 12, no. 11, Nov. 1998, pp. 1661–72. Pubmed, doi:10.1210/mend.12.11.0192.

Published In

Mol Endocrinol

DOI

ISSN

0888-8809

Publication Date

November 1998

Volume

12

Issue

11

Start / End Page

1661 / 1672

Location

United States

Related Subject Headings

  • Structure-Activity Relationship
  • Repetitive Sequences, Amino Acid
  • Recombinant Fusion Proteins
  • Receptor, IGF Type 2
  • Protein Structure, Tertiary
  • Protein Binding
  • Insulin-Like Growth Factor II
  • Humans
  • Genes, Synthetic
  • Endocrinology & Metabolism