title: UDP-glucose pyrophosphorylase from potato tuber: purification and characterization.
abstract: UDP-glucose pyrophosphorylase from potato tuber was purified 243-fold to a nearly homogeneous state with a recovery of 30%. The purified enzyme utilized UDP-glucose, but not ADP-glucose, as the substrate, and was not activated by 3-phosphoglyceric acid. Product inhibition studies revealed the sequential binding of UDP-glucose and MgPPi and the sequential release of glucose-1-phosphate and MgUTP, in this order. Analyses of the effects of Mg2+ on the enzyme activity suggest that the MgPPi and MgUTP complexes are the actual substrates for the enzyme reaction, and that free UTP acts as an inhibitor. The enzyme exists probably as the monomer of an approximately 50-kDa polypeptide with a blocked amino terminus. For structural comparison, 29 peptides isolated from a tryptic digest of the S-carboxymethylated enzyme were sequenced. The results show that the potato tuber enzyme is homologous to UDP-glucose pyrophosphorylase from slime mold, but not to ADP-glucose pyrophosphorylase from Escherichia coli, and provide structural evidence that UDP-glucose and ADP-glucose pyrophosphorylase are two different protein entities.
authors.affiliation: Institute of Scientific and Industrial Research, Osaka University.
authors.forename: K
authors.initials: K
authors.lastname: Nakano
authors.forename: Y
authors.initials: Y
authors.lastname: Omura
authors.forename: M
authors.initials: M
authors.lastname: Tagaya
authors.forename: T
authors.initials: T
authors.lastname: Fukui
mesh_terms: D000595:Amino Acid Sequence; D007700:Kinetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009713:Nucleotidyltransferases / Q000302:isolation & purification*; D010944:Plants / Q000201:enzymology*; D012689:Sequence Homology, Nucleic Acid; D011198:Solanum tuberosum / Q000032:analysis; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000302:isolation & purification* / Q000378:metabolism; D014532:Uridine Diphosphate Glucose / Q000378:metabolism
chemical_list: D009713:Nucleotidyltransferases; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase; D014532:Uridine Diphosphate Glucose