Distinct subcellular localisations of the putative inositol 1,3,4,5-tetrakisphosphate receptors GAP1IP4BP and GAP1m result from the GAP1IP4BP PH domain directing plasma membrane targeting

Curr Biol. 1997 Dec 1;7(12):1007-10. doi: 10.1016/s0960-9822(06)00423-4.

Abstract

Inositol 1,3,4,5-tetrakisphosphate (IP4), is a ubiquitous inositol phosphate that has been suggested to function as a second messenger. Recently, we purified and cloned a putative IP4 receptor, termed GAP1(IP4BP)[1], which is also a member of the GAP1 family of GTPase-activating proteins for the Ras family of GTPases. A homologue of GAP1(IP4BP), called GAP1(m), has been identified [2] and here we describe the cloning of a GAP1(m) cDNA from a human circulating-blood cDNA library. We found that a deletion mutant of GAP1(m), in which the putative phospholipid-binding domains (C2A and C2B) have been removed, binds to IP4 with a similar affinity and specificity to that of the corresponding GAP1(IP4BP) mutant. Expression studies of the proteins in either COS-7 or HeLa cells showed that, whereas GAP1(IP4BP) is located solely at the plasma membrane, GAP1(m) seems to have a distinct perinuclear localisation. By mutational analysis, we have shown that the contrast in subcellular distribution of these two closely related proteins may be a function of their respective pleckstrin homology (PH) domains. This difference in localisation has fundamental significance for our understanding of the second messenger functions of IP4.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Blood Proteins / chemistry
  • COS Cells
  • Cell Membrane / metabolism
  • HeLa Cells
  • Humans
  • Inositol Phosphates / metabolism*
  • Phosphoproteins*
  • Proteins / chemistry
  • Proteins / genetics
  • Proteins / metabolism*
  • Receptors, Cytoplasmic and Nuclear / chemistry
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sequence Deletion
  • Structure-Activity Relationship
  • Subcellular Fractions / metabolism
  • ras GTPase-Activating Proteins*

Substances

  • Blood Proteins
  • Inositol Phosphates
  • Phosphoproteins
  • Proteins
  • RASA2 protein, human
  • Receptors, Cytoplasmic and Nuclear
  • Recombinant Fusion Proteins
  • inositol-1,3,4,5-tetrakisphosphate receptor
  • platelet protein P47
  • ras GTPase-Activating Proteins
  • inositol-1,3,4,5-tetrakisphosphate