Identification of arfophilin, a target protein for GTP-bound class II ADP-ribosylation factors

Ok Ho Shin, Annette H. Ross, Irene Mihai, John H. Exton

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Yeast two-hybrid screening of a human kidney cDNA library using the GTP- bound form of a class II ADP-ribosylation factor (ARF5) identified a novel ARF5-binding protein with a calculated molecular mass of 82.4 kDa, which was named arfophilin. Northern hybridization analysis showed high level arfophilin mRNA expression in human heart and skeletal muscle. Arfophilin bound only to the active, GTP-bound form of ARF5 and did not bind to GTP- ARF3, which is a class I ARF. The N terminus of ARF5 (1-17 amino acids) was essential for binding to arfophilin. The GTP-bound form of ARF5 with amino acid residues in the N terminus mutated to those in ARF4 (another class II ARF) also bound to arfophilin, suggesting it is a target protein for GTP- bound forms of class II ARFs. The binding site for ARF on arfophilin was localized to the C terminus (residues 612-756), which contains putative coiled-coil structures. Recombinant arfophilin overexpressed in CHO-K1 cells was localized in the cytosol and translocated to a membrane fraction in association with GTP-bound ARF5. ARF5 containing the N terminus of ARF3 did not promote translocation indicating that class II ARFs are specific carriers for arfophilin.

Original languageEnglish (US)
Pages (from-to)36609-36615
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number51
DOIs
StatePublished - Dec 17 1999
Externally publishedYes

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ADP-Ribosylation Factors
Guanosine Triphosphate
Proteins
Amino Acids
Essential Amino Acids
arfophilin
CHO Cells
Molecular mass
Gene Library
Cytosol
Yeast
Muscle
Myocardium
Carrier Proteins
Screening
Skeletal Muscle
Yeasts
Binding Sites
Membranes
Kidney

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of arfophilin, a target protein for GTP-bound class II ADP-ribosylation factors. / Shin, Ok Ho; Ross, Annette H.; Mihai, Irene; Exton, John H.

In: Journal of Biological Chemistry, Vol. 274, No. 51, 17.12.1999, p. 36609-36615.

Research output: Contribution to journalArticle

Shin, Ok Ho ; Ross, Annette H. ; Mihai, Irene ; Exton, John H. / Identification of arfophilin, a target protein for GTP-bound class II ADP-ribosylation factors. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 51. pp. 36609-36615.
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