High-affinity binding of a FYVE domain to phosphatidylinositol 3-phosphate requires intact phospholipid but not FYVE domain oligomerization

Biochemistry. 2001 Jul 24;40(29):8581-7. doi: 10.1021/bi010425d.

Abstract

FYVE domains are small zinc-finger-like domains found in many proteins that are involved in regulating membrane traffic and have been shown to bind specifically to phosphatidylinositol 3-phosphate (PtdIns-3-P). FYVE domains are thought to recruit PtdIns-3-P effectors to endosomal locations in vivo, where these effectors participate in controlling endosomal maturation and vacuolar protein sorting. We have compared the characteristics of PtdIns-3-P binding by the FYVE domain from Hrs-1 (the hepatocyte growth factor-regulated tyrosine kinase substrate) with those of specific phosphoinositide binding by Pleckstrin homology (PH) domains. Like certain PH domains (such as that from phospholipase C-delta(1)), the Hrs-1 FYVE domain specifically recognizes a single phosphoinositide. However, while phosphoinositide binding by highly specific PH domains is driven almost exclusively by interactions with the lipid headgroup, this is not true for the Hrs-1 FYVE domain. The phospholipase C-delta(1) PH domain shows a 10-fold preference for binding isolated headgroup over its preferred lipid (phosphatidylinositol 4,5-bisphosphate) in a membrane, while the Hrs-1 FYVE domain greatly prefers (more than 50-fold) intact lipid in a bilayer over the isolated headgroup (inositol 1,3-bisphosphate). By contrast with reports for certain PH domains, we find that this preference for membrane binding over interaction with soluble lipid headgroups does not require FYVE domain oligomerization.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Binding, Competitive / genetics
  • Blood Proteins / genetics
  • Blood Proteins / metabolism
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Cation Transport Proteins*
  • Glutathione Transferase / genetics
  • Guanine Nucleotide Exchange Factors
  • HeLa Cells
  • Humans
  • Liposomes / metabolism
  • Monosaccharide Transport Proteins*
  • Phosphatidylinositol Phosphates / metabolism*
  • Phospholipids / metabolism*
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Protein Binding / genetics
  • Protein Structure, Tertiary / genetics
  • Proteins / genetics
  • Proteins / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Symporters*
  • Zinc Fingers* / genetics

Substances

  • Blood Proteins
  • Carrier Proteins
  • Cation Transport Proteins
  • FGD1 protein, human
  • Guanine Nucleotide Exchange Factors
  • Liposomes
  • Monosaccharide Transport Proteins
  • Phosphatidylinositol Phosphates
  • Phospholipids
  • Phosphoproteins
  • Proteins
  • RSC1A1 protein, human
  • Recombinant Fusion Proteins
  • Symporters
  • phosphatidylinositol 3-phosphate
  • platelet protein P47
  • Glutathione Transferase