Importance of N-terminal regions of G protein alpha subunits for the activation of phospholipase C in Xenopus oocytes.
The alpha subunits of Gq family G proteins, GL1 alpha and GL2 alpha, are bovine homologues of mouse G14 alpha and G11 alpha, respectively, and are closely related to each other. When expressed in Xenopus oocytes together with metabotropic glutamate receptors, GL2 alpha activates endogenous phospholipase C (PLCx) in response to glutamate stimulation, whereas GL1 alpha inhibits the activation of PLCx. By examining the properties of 10 chimeras between GL1 alpha and GL2 alpha and their mutants, we tried to identify the regions on the G alpha proteins that are important for the activation of PLCx. The results indicated that a necessary (but not sufficient) condition for a chimeric G alpha protein to be able to clearly activate PLCx was that its N-terminal quarter portion should be derived from GL2 alpha. No correlation was found between the origin (GL1 alpha or GL2 alpha) of C-terminal regions of the chimeras and the ability of chimeras to activate PLCx. One of the chimeras is different from GL2 alpha at only four amino acid residues in the N-terminal region, and yet it could not activate PLCx. When one of the four residues, Ser-59, in the chimera was mutated back to Ala as in the original GL2 alpha, the resulting mutant became capable of activating PLCx. This residue is localized in the midst of the N-terminal linker connecting the two major domains in the G alpha proteins. These results indicate that Ala-59 is critical for the activation of PLCx, and that the linker may play important roles in determining functions of G alpha proteins.
Nakamura
K
K
Department of Molecular Biology, Graduate School of Medical Science, Kyushu University, Fukuoka.
Nukada
T
T
Imai
K
K
Sugiyama
H
H
D000818:Animals; D002417:Cattle; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000737:chemistry*; D051379:Mice; D009865:Oocytes / Q000201:enzymology*; D011487:Protein Conformation; D017470:Receptors, Glutamate / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D010738:Type C Phospholipases / Q000378:metabolism*; D014981:Xenopus
D017470:Receptors, Glutamate; D011993:Recombinant Fusion Proteins; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins