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• W1974029229 endingPage "1672" @default.
• W1974029229 startingPage "1661" @default.
• W1974029229 abstract "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 ∼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." @default.
• W1974029229 created "2016-06-24" @default.
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• W1974029229 date "1998-11-01" @default.
• W1974029229 modified "2023-10-18" @default.
• W1974029229 title "An Insulin-Like Growth Factor II (IGF-II) Affinity-Enhancing Domain Localized within Extracytoplasmic Repeat 13 of the IGF-II/Mannose 6-Phosphate Receptor" @default.
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• W1974029229 doi "https://doi.org/10.1210/mend.12.11.0192" @default.
• W1974029229 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/9817593" @default.
• W1974029229 hasPublicationYear "1998" @default.
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