Arfophilin is a common target of both class II and class III ADP-ribosylation factors

O. H. Shin, A. D. Couvillon, J. H john Exton

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Arfophilin was first identified as a target protein for GTP-ARF5. The N-terminus of ARF5 (armno acids 2-17), which is distinct from that of class I or class III ARFs, is essential for binding to the C-terminus of arfophilin (amino acids 612-756). This study using GST fusion proteins in pulldown experiments in CHO-K1 cell lysates showed that, unexpectedly, ARF6 also bound to full-length arfophilin or the C-terminus of arfophilin (amino acids 612-756) in a GTP-dependent manner. Studies with ARF1/ ARF6 chimeras further showed that the amino acid sequence of residues 37-80 of ARF6, which is different from the corresponding sequences in class I and class II ARFs, was essential for binding to arfophilin. Both GTP-ARF5 and GTP-ARF6 bound to arfophilin in CHO-K1 cell lysates, while GTP-ARF1 did not bind. In contrast, all three forms of ARF bound to arfaptin 2, with ARF1 showing the strongest binding. Yeast two-hybrid studies with wild-type, dominant negative, and constitutively active forms of ARF1, -5, and -6 and with ARF1/ARF6 chimeras confirmed these results, except that constitutively active ARF6 was autoactivating. Our findings suggest that both class II and III ARFs may influence the same cellular pathways through arfophilin as a common downstream effector.

Original languageEnglish (US)
Pages (from-to)10846-10852
Number of pages7
JournalBiochemistry
Volume40
Issue number36
DOIs
StatePublished - Sep 11 2001
Externally publishedYes

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ADP-Ribosylation Factors
Guanosine Triphosphate
CHO Cells
Amino Acids
arfophilin
Yeast
Amino Acid Sequence
Proteins
Fusion reactions
Yeasts
Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Arfophilin is a common target of both class II and class III ADP-ribosylation factors. / Shin, O. H.; Couvillon, A. D.; Exton, J. H john.

In: Biochemistry, Vol. 40, No. 36, 11.09.2001, p. 10846-10852.

Research output: Contribution to journalArticle

Shin, O. H. ; Couvillon, A. D. ; Exton, J. H john. / Arfophilin is a common target of both class II and class III ADP-ribosylation factors. In: Biochemistry. 2001 ; Vol. 40, No. 36. pp. 10846-10852.
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