Palmitoylation but not the extreme amino-terminus of Gq alpha is required for coupling to the NK2 receptor

FEBS Lett. 1994 Nov 7;354(2):195-9. doi: 10.1016/0014-5793(94)01101-x.

Abstract

Gq alpha and G11 alpha differ from other G protein alpha subunits in that they have unique, conserved 6 residue amino-terminal extensions. Wild-type and amino-terminal mutants of Gq alpha expressed in COS cells were analyzed for their ability to functionally couple with co-expressed neurokinin NK2 receptor. Wild-type, T2A and delta 2-7 Gq alpha were able to stimulate agonist driven phospholipase C (PLC) activity in identical manners. Other activities of these two amino-terminal mutants including aluminum fluoride stimulated PLC activity, palmitoylation, interaction with G beta gamma subunits and GTP gamma S-induced trypsin resistance are also similar to the wild-type alpha subunit. This demonstrates that the NK2 receptor is able to functionally interact with the alpha subunit of Gq and that the first seven amino-acids of Gq alpha are not required for any of the alpha subunit functions tested. In contrast to the T2A and delta 2-7 mutants, a C9,10A Gq alpha mutant was not able to couple to either the NK2 receptor or PLC, as assessed by high-affinity agonist binding and activation of PLC either in intact cells or in vitro. The C9,10A protein was able to assume a GTP gamma S-induced trypsin-resistant conformation and partitioned primarily to the pelletable fraction in a manner similar to the wild-type protein. However, it was not labeled with [3H]palmitic acid. This suggests that blocking palmitoylation at the amino-terminus of Gq alpha results in a loss of functional activity which reflects an inability to interact with both the receptor and downstream signaling targets.

MeSH terms

  • Aluminum Compounds / pharmacology
  • Animals
  • Cell Line
  • Cytidine Diphosphate Diglycerides / metabolism
  • Detergents
  • Enzyme Activation
  • Fluorides / pharmacology
  • GTP-Binding Proteins / chemistry
  • GTP-Binding Proteins / genetics
  • GTP-Binding Proteins / metabolism*
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology
  • Mutagenesis
  • Palmitic Acid
  • Palmitic Acids / metabolism*
  • Plasmids
  • Receptors, Neurokinin-2 / genetics
  • Receptors, Neurokinin-2 / metabolism*
  • Recombinant Proteins / metabolism
  • Solubility
  • Structure-Activity Relationship
  • Transfection
  • Type C Phospholipases / metabolism

Substances

  • Aluminum Compounds
  • Cytidine Diphosphate Diglycerides
  • Detergents
  • Palmitic Acids
  • Receptors, Neurokinin-2
  • Recombinant Proteins
  • Palmitic Acid
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Type C Phospholipases
  • GTP-Binding Proteins
  • Fluorides
  • aluminum fluoride