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Identification of a region at the N-terminus of phospholipase C-beta 3 that interacts with G protein beta gamma subunits.

Publication ,  Journal Article
Barr, AJ; Ali, H; Haribabu, B; Snyderman, R; Smrcka, AV
Published in: Biochemistry
February 22, 2000

Members of the phospholipase C-beta (PLC-beta) family of proteins are activated either by G alpha or G beta gamma subunits of heterotrimeric G proteins. To define specific regions of PLC-beta 3 that are involved in binding and activation by G beta gamma, a series of fragments of PLC-beta 3 as glutathione-S-transferase (GST) fusion proteins were produced. A fragment encompassing the N-terminal pleckstrin homology (PH) domain and downstream sequence (GST-N) bound to G protein beta 1 gamma 2 in an in vitro binding assay, and binding was inhibited by G protein alpha subunit, G alpha i1. This PLC-beta 3 fragment also inhibited G beta gamma-stimulated PLC-beta activity in a reconstitution system, while having no significant effect on G alpha q-stimulated PLC-beta 3 activity. The N-terminal G beta gamma binding region was delineated further to the first 180 amino acids, and the sequence Asn150-Ser180, just distal to the PH domain, was found to be required for the interaction. Mutation of basic residues 154Arg, 155Lys, 159Lys, and 161Lys to Glu within this region reduced G beta gamma binding affinity and specifically reduced the EC50 for G beta gamma-dependent activation of the mutant enzyme 3-fold. Basal activity and G alpha q-dependent activation of the enzyme were unaffected by the mutations. While these basic residues may not directly mediate the interaction with G beta gamma, the data provide evidence for an N-terminal G beta gamma binding region of PLC-beta 3 that is involved in activation of the enzyme.

Duke Scholars

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

February 22, 2000

Volume

39

Issue

7

Start / End Page

1800 / 1806

Location

United States

Related Subject Headings

  • Type C Phospholipases
  • Recombinant Fusion Proteins
  • Proto-Oncogene Proteins
  • Protein Binding
  • Point Mutation
  • Phospholipase C beta
  • Peptide Fragments
  • Molecular Sequence Data
  • Isoenzymes
  • Humans
 

Citation

APA
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MLA
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Barr, A. J., Ali, H., Haribabu, B., Snyderman, R., & Smrcka, A. V. (2000). Identification of a region at the N-terminus of phospholipase C-beta 3 that interacts with G protein beta gamma subunits. Biochemistry, 39(7), 1800–1806. https://doi.org/10.1021/bi992021f
Barr, A. J., H. Ali, B. Haribabu, R. Snyderman, and A. V. Smrcka. “Identification of a region at the N-terminus of phospholipase C-beta 3 that interacts with G protein beta gamma subunits.Biochemistry 39, no. 7 (February 22, 2000): 1800–1806. https://doi.org/10.1021/bi992021f.
Barr AJ, Ali H, Haribabu B, Snyderman R, Smrcka AV. Identification of a region at the N-terminus of phospholipase C-beta 3 that interacts with G protein beta gamma subunits. Biochemistry. 2000 Feb 22;39(7):1800–6.
Barr, A. J., et al. “Identification of a region at the N-terminus of phospholipase C-beta 3 that interacts with G protein beta gamma subunits.Biochemistry, vol. 39, no. 7, Feb. 2000, pp. 1800–06. Pubmed, doi:10.1021/bi992021f.
Barr AJ, Ali H, Haribabu B, Snyderman R, Smrcka AV. Identification of a region at the N-terminus of phospholipase C-beta 3 that interacts with G protein beta gamma subunits. Biochemistry. 2000 Feb 22;39(7):1800–1806.
Journal cover image

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

February 22, 2000

Volume

39

Issue

7

Start / End Page

1800 / 1806

Location

United States

Related Subject Headings

  • Type C Phospholipases
  • Recombinant Fusion Proteins
  • Proto-Oncogene Proteins
  • Protein Binding
  • Point Mutation
  • Phospholipase C beta
  • Peptide Fragments
  • Molecular Sequence Data
  • Isoenzymes
  • Humans