Selective modification by transglutaminase of a glutamine side chain in the hinge region of the histidine-388----glutamine mutant of yeast phosphoglycerate kinase.
The transglutaminase-catalysed incorporation of putrescine and monodansylcadaverine into yeast phosphoglycerate kinase has been studied. There is little incorporation of the amines into wild-type enzyme, but nearly stoichiometric incorporation into the histidine-388----glutamine mutant enzyme. C.d. studies show that the overall structure of the mutant enzyme is very similar to that of the wild-type enzyme. Incorporation of the amines into the mutant enzyme causes no significant change in its activity. Glutamine-388 was shown, by isolation and sequencing of the modified peptide, to be the site of incorporation of monodansylcadaverine into the mutant enzyme. The specificity of the transglutaminase reaction is discussed in the light of available data.
Coussons
P J
PJ
Department of Biological and Molecular Sciences, University of Stiriling, Scotland, U.K.
Kelly
S M
SM
Price
N C
NC
Johnson
C M
CM
Smith
B
B
Sawyer
L
L
D000595:Amino Acid Sequence; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D005973:Glutamine / Q000235:genetics*; D006639:Histidine / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010449:Peptide Mapping; D010735:Phosphoglycerate Kinase / Q000235:genetics* / Q000378:metabolism; D011700:Putrescine / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D013056:Spectrophotometry, Ultraviolet; D011503:Transglutaminases; D014357:Trypsin
D005973:Glutamine; D006639:Histidine; D011503:Transglutaminases; D010735:Phosphoglycerate Kinase; D014357:Trypsin; D011700:Putrescine