Specificity of G protein alpha-gamma subunit interactions. N-terminal 15 amino acids of gamma subunit specifies interaction with alpha subunit.
The existence of multiple alpha, beta, and gamma subunits raises questions regarding the assembly of particular G proteins. Based on the results of a previous study (Rahmatullah, M., and Robishaw, J. D. (1994) J. Biol. Chem. 269, 3574-3580), we hypothesized that the assembly of G proteins may be determined by the interactions of the more structurally diverse alpha and gamma subunits. This hypothesis was confirmed in the present study by showing striking differences in the abilities of the gamma 1 and gamma 2 subunits to interact with the alpha o subunit. Chimeras of the gamma 1 and gamma 2 subunits were used to delineate which region is responsible. Support for the importance of the N-terminal region of the gamma subunit comes from our observations that 1) the gamma 2 subunit and the gamma 211 chimera bound strongly to the alpha o-agarose matrix, but the gamma 1 subunit and the gamma 112 chimera bound weakly, if at all; 2) an N-terminal peptide made to the gamma 2 subunit blocked the binding of the gamma 211 chimera to the alpha o-agarose matrix; 3) both the gamma 211 chimera and the N-terminal peptide were able to partially protect the alpha o subunit against tryptic cleavage; and 4) the gamma 211 chimera, but not the gamma 112 chimera, supported ADP-ribosylation of the alpha o subunit.
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania 17822.
M
M
Rahmatullah
R
R
Ginnan
J D
JD
Robishaw
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D015151:Immunoblotting; D046911:Macromolecular Substances; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D037342:Pertussis Toxin; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera; D014162:Transfection; D014357:Trypsin; D010566:Virulence Factors, Bordetella / Q000378:metabolism / Q000494:pharmacology
D046911:Macromolecular Substances; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D037342:Pertussis Toxin; D014357:Trypsin; D019204:GTP-Binding Proteins