The ADP-glucose pyrophosphorylase from Escherichia coli comprises two tightly bound distinct domains.
Computational analysis of ADP-glucose pyrophosphorylases predicts a fold with two domains. Co-expression of two polypeptides comprising residues 1-323 and 328-431 from the Escherichia coli ADP-glucose pyrophosphorylase yielded an enzyme form as active as the wild type. The only difference from the wild type was a slightly modified affinity for allosteric effectors. The two polypeptides could not be separated by chromatographic procedures. Separate expression of these polypeptides produced inactive unstable forms. All these results indicated that the ADP-glucose pyrophosphorylase comprises two domains with a strong interaction between them. That interaction is important for allosteric properties and structural stability.
Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824, USA.
Clarisa M
CM
Bejar
Miguel A
MA
Ballicora
Diego F
DF
Gómez-Casati
Alberto A
AA
Iglesias
Jack
J
Preiss
D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D001483:Base Sequence; D020134:Catalytic Domain; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D051599:Glucose-1-Phosphate Adenylyltransferase; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase