title: ADPG formation by the ADP-specific cleavage of sucrose-reassessment of sucrose synthase.
abstract: The standardized enzyme coupling method for assaying sucrose synthase activities in the direction of sucrose cleavage was reexamined using enzyme preparations from cultured cells of sycamore (Acer pseudoplatanus L.) and spinach leaves (Spinacea oleracea). Both ATP and Tris, commonly utilized in assay systems to measure sucrose synthase, were found to inhibit non-competitively the ADPG-synthesizing activities of the enzyme. Upon substituting ATP by either GTP or UTP, and Tris by HEPES, we found that the sucrose synthase is capable of producing ADPG effectively, recognizing ADP as the principal substrate (Km = 5.3 microM (sycamore) and 16.8 microM (spinach]. The Vmax value for the synthesis of ADPG clearly surpasses the Vmax observed for the synthesis of UDPG by the enzyme. It was found that UDP is not inhibitory on the synthesis of ADPG by SS, which behaves allosterically with respect to the concentration level of sucrose.
authors.affiliation: Research Institute for Biochemical Regulation, School of Agriculture, Nagoya University, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Pozueta-Romero
authors.forename: J
authors.initials: J
authors.lastname: Yamaguchi
authors.forename: T
authors.initials: T
authors.lastname: Akazawa
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism*; D000245:Adenosine Diphosphate Glucose / Q000096:biosynthesis*; D000255:Adenosine Triphosphate / Q000494:pharmacology; D002384:Catalysis; D005964:Glucosyltransferases / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D010944:Plants / Q000201:enzymology; D006180:Proton-Translocating ATPases / Q000378:metabolism; D013379:Substrate Specificity / Q000187:drug effects; D013395:Sucrose / Q000378:metabolism*; D014325:Tromethamine; D043324:beta-Fructofuranosidase
chemical_list: D014325:Tromethamine; D000245:Adenosine Diphosphate Glucose; D013395:Sucrose; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D005964:Glucosyltransferases; C022699:sucrose synthase; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D006180:Proton-Translocating ATPases