Inositol 1,3,4,5-tetrakisphosphate and inositol hexakisphosphate receptor proteins: Isolation and characterization from rat brain

Anne B. Theibert, Virginia A. Estevez, Christopher D. Ferris, Sonye K. Danoff, Roxanne K. Barrow, Glenn D. Prestwich, Solomon H. Snyder

Research output: Contribution to journalArticle

Abstract

High-affinity, membrane-associated inositol 1,3,4,5-tetrakisphosphate (IP4) and inositol hexakisphosphate (IP6) binding proteins were solubilized and isolated utilizing a heparin-agarose resin followed by an IP4 affinity resin. The IP6 receptor comprises a protein complex of 115-, 105-, and 50-kDa subunits, all of which comigrate under native conditions. The Kd of the receptor for IP6 is 12 nM, whereas inositol 1,3,4,5,6-pentakisphosphate (IP5), IP4, and inositol 1,4,5-trisphosphate (IP3) are 50%, 30%, and 15%, respectively, as potent. Two protein complexes copurify with the IP4 receptor fraction. A 182/123-kDa complex elutes first from the affinity column followed by a 174/84-kDa protein complex, which elutes at higher salt. Both complexes show high affinity for IP4 (Kd = 3-4 nM). IP5, IP6, and IP3 display approximately 25%, 10%, and 0.1%, respectively, the affinity of IP4. Ligand binding to IP6 and IP4 receptors is inhibited 50% by heparin at 0.1 μg/ml. IP4 receptor proteins are stoichiometrically phospborylated by cyclic AMP-dependent protein kinase and protein kinase C, whereas negligible phosphorylation is observed for the IP6 receptor. (.

Original languageEnglish (US)
Pages (from-to)3165-3169
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number8
DOIs
StatePublished - 1991

Keywords

  • Cyclic AMP-dependent protein kinase
  • Inositol phosphates
  • Protein kinase C

ASJC Scopus subject areas

  • General

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