Determination of interaction sites on the small G protein RhoA for phospholipase D

Chang Dae Bae, Do Sik Min, Ian N. Fleming, John H. Exton

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Phespholipase D (PLD) has been identified as a target of small G proteins of the Rho family. The present study was directed at defining the interaction sites of RhoA with rat brain PLD in vitro using chimeric proteins between RhoA and Ha-Has or Cdc42Hs and point mutations. The switch I region of RhoA, which is the common effector domain of Ras-like G proteins, was a crucial interaction site for PLD. Mutations in conserved amino acids (Tyr34, Thr37, Phe39) totally abolished PLD activation, while mutations in Val38 or Tyr42 caused partial loss. Two additional sites were responsible for the differential PLD activation ability between RhoA and Cdc42Hs. Changing Asp76 in the switch II region of RhoA to the corresponding amino acid in Cdc42Hs led to partial loss of PLD activation. A chimeric protein with the N-terminal third of Cdc42Hs changed to RhoA showed enhanced PLD activation. Analysis of other Rho/Ha-Ras chimeric proteins and mutations indicated that Gln52 adjacent to the switch II region is responsible for this gain of function. In conclusion, the present study shows that conserved amino acids in the switch I region of RhoA are major PLD interaction sites and that residues in the switch H and internal regions are responsible for the differential activation of PLD by RhoA and Cdc42Hs.

Original languageEnglish
Pages (from-to)11596-11604
Number of pages9
JournalJournal of Biological Chemistry
Volume273
Issue number19
DOIs
Publication statusPublished - 1998 May 8

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Phospholipase D
Monomeric GTP-Binding Proteins
Chemical activation
Switches
Amino Acids
Mutation
rhoA GTP-Binding Protein
ras Proteins
GTP-Binding Proteins
Point Mutation
Proteins
Brain
Rats

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Bae, Chang Dae ; Min, Do Sik ; Fleming, Ian N. ; Exton, John H. / Determination of interaction sites on the small G protein RhoA for phospholipase D. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 19. pp. 11596-11604.
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Determination of interaction sites on the small G protein RhoA for phospholipase D. / Bae, Chang Dae; Min, Do Sik; Fleming, Ian N.; Exton, John H.

In: Journal of Biological Chemistry, Vol. 273, No. 19, 08.05.1998, p. 11596-11604.

Research output: Contribution to journalArticle

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