Characterization of a novel phosphatidylinositol 3-phosphate-binding protein containing two FYVE fingers in tandem that is targeted to the Golgi

P C Cheung, L Trinkle-Mulcahy, P Cohen, J M Lucocq

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

We have identified a novel protein of predicted molecular mass 40 kDa that contains two FYVE domains in tandem and has therefore been named TAFF1 (TAndem FYVE Fingers-1). The protein is expressed predominantly in heart and binds to PtdIns3P specifically, even though the FYVE domains in TAFF1 lacks the first Arg of the consensus sequence R(K/R)HHCR, critical for the PtdIns3P binding of other FYVE domains identified so far. The first Arg is replaced by a Thr and Ser in the N-terminal and C-terminal FYVE domains of TAFF1 respectively. Mutational analysis indicates that both FYVE domains are required for high affinity binding to PtdIns3P. Cell localization studies using a green fluorescent protein fusion show that TAFF1 is localized to the Golgi, and that the Golgi targeting sequence is located within the N-terminal 187 residues and not in either FYVE domain.
Original languageEnglish
Pages (from-to)113-21
Number of pages9
JournalBiochemical Journal
Volume355
Issue numberPt 1
Publication statusPublished - 1 Apr 2001

Keywords

  • Amino Acid Sequence
  • Base Sequence
  • Blotting, Northern
  • Carrier Proteins
  • DNA, Complementary
  • Golgi Apparatus
  • Green Fluorescent Proteins
  • Humans
  • Luminescent Proteins
  • Microscopy, Immunoelectron
  • Molecular Sequence Data
  • Recombinant Fusion Proteins
  • Sequence Homology, Amino Acid
  • Subcellular Fractions
  • Two-Hybrid System Techniques

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