A (His)6-tagged recombinant barley (Hordeum vulgare L.) endosperm ADP-glucose pyrophosphorylase expressed in the baculovirus-insect cell system is insensitive to allosteric regulation by 3-phosphoglycerate and inorganic phosphate.
ADP-glucose pyrophosphorylase from photosynthetic tissue is allosterically regulated by 3-phosphoglycerate and inorganic phosphate. In contrast, data from our laboratory indicated that the major AGPase from barley seeds is insensitive to these regulators. Verification of this conclusion has, however, been hindered by the proteolytic degradation of the enzyme from seeds. This report characterizes the barley seed AGPase expressed in the baculovirus-insect cell system, confirming that lack of allosteric regulation by 3-PGA/Pi is an intrinsic property of the enzyme. Purification of the enzyme was by Ni2+-NTA agarose chromatography using a (His)6 tag attached to the N-terminus of the small AGPase subunit.
Rudi
H
H
Department of Biotechnological Sciences, Agricultural University of Norway, As.
Doan
D N
DN
Olsen
O A
OA
D000494:Allosteric Regulation / Q000187:drug effects; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics; D051599:Glucose-1-Phosphate Adenylyltransferase; D005988:Glyceric Acids / Q000494:pharmacology*; D006639:Histidine; D001467:Hordeum / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010455:Peptides / Q000235:genetics; D010710:Phosphates / Q000494:pharmacology*; D010802:Phylogeny; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera
D005988:Glyceric Acids; D010455:Peptides; D010710:Phosphates; D011993:Recombinant Fusion Proteins; C033223:polyhistidine; D006639:Histidine; C005156:3-phosphoglycerate; D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase