title: Relative turnover numbers of maize endosperm and potato tuber ADP-glucose pyrophosphorylases in the absence and presence of 3-phosphoglyceric acid.
abstract: Adenosine diphosphate glucose pyrophosphorylase (AGPase; EC 2.7.7.27) synthesizes the starch precursor, ADP-glucose. It is a rate-limiting enzyme in starch biosynthesis and its activation by 3-phosphoglyceric acid (3PGA) and/or inhibition by inorganic phosphate (Pi) are believed to be physiologically important. Leaf, tuber and cereal embryo AGPases are highly sensitive to these effectors, whereas endosperm AGPases are much less responsive. Two hypotheses can explain the 3PGA activation differences. Compared to leaf AGPases, endosperm AGPases (i) lack the marked ability to be activated by 3PGA or (ii) they are less dependent on 3PGA for activity. The absence of purified preparations has heretofore negated answering this question. To resolve this issue, heterotetrameric maize ( Zea mays L.) endosperm and potato ( Solanum tuberosum L.) tuber AGPases expressed in Escherichia coli were isolated and the relative amounts of enzyme protein were measured by reaction to antibodies against a motif resident in both small subunits. Resulting reaction rates of both AGPases are comparable in the presence but not in the absence of 3PGA when expressed on an active-protein basis. We also placed the potato tuber UpReg1 mutation into the maize AGPase. This mutation greatly enhances 3PGA sensitivity of the potato AGPase but it has little effect on the maize AGPase. Thirdly, lysines known to bind 3PGA in potato tuber AGPase, but missing from the maize endosperm AGPase, were introduced into the maize enzyme. These had minimal effect on maize endosperm activity. In conclusion, the maize endosperm AGPase is not nearly as dependent on 3PGA for activity as is the potato tuber AGPase.
authors.affiliation: Program in Plant Molecular and Cellular Biology, Horticultural Sciences, University of Florida, Gainesville 32611, USA.
authors.forename: Brian T
authors.initials: BT
authors.lastname: Burger
authors.forename: Joanna M
authors.initials: JM
authors.lastname: Cross
authors.forename: Janine R
authors.initials: JR
authors.lastname: Shaw
authors.forename: Joel R
authors.initials: JR
authors.lastname: Caren
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Greene
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Okita
authors.forename: L Curtis
authors.initials: LC
authors.lastname: Hannah
mesh_terms: D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D051599:Glucose-1-Phosphate Adenylyltransferase; D005988:Glyceric Acids / Q000494:pharmacology*; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010710:Phosphates / Q000494:pharmacology; D018515:Plant Leaves / Q000187:drug effects / Q000201:enzymology; D018547:Plant Stems / Q000187:drug effects / Q000201:enzymology; D012639:Seeds / Q000187:drug effects / Q000201:enzymology; D011198:Solanum tuberosum / Q000201:enzymology* / Q000235:genetics; D003313:Zea mays / Q000201:enzymology* / Q000235:genetics
chemical_list: D004791:Enzyme Inhibitors; D005988:Glyceric Acids; D010710:Phosphates; C005156:3-phosphoglycerate; D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase