Studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. I. Characterization and properties of the enzyme
Phosphorylase II from potato tuber has been subjected to several purification procedures in order to: (a) separate the unprimed activity from the primed one coexisting in the enzymatic preparation, and (b) eliminate endogenous glucan and/or glycoproteins. Large-scale analytical nondenaturing gel ele...
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todo:paper_00039861_v212_n2_p525_Sivak2023-10-03T13:56:56Z Studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. I. Characterization and properties of the enzyme Sivak, M.N. Tandecarz, J.S. Cardini, C.E. isoenzyme phosphorylase article enzyme specificity enzymology heat isolation and purification kinetics metabolism molecular weight plant Heat Isoenzymes Kinetics Molecular Weight Phosphorylases Plants Substrate Specificity Support, Non-U.S. Gov't Phosphorylase II from potato tuber has been subjected to several purification procedures in order to: (a) separate the unprimed activity from the primed one coexisting in the enzymatic preparation, and (b) eliminate endogenous glucan and/or glycoproteins. Large-scale analytical nondenaturing gel electrophoresis and affinity chromatography on concanavalin A-Sepharose 4B succeeded in removing endogenous glycoproteins without any effect on the unprimed activity. In addition, phosphorolysis or glucoamylase treatment of the enzymatic preparation did not abolish the phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. However, no separation between both activities, primed and unprimed, could be achieved either by the above-mentioned methods or by sucrose density gradient centrifugation. Based on sodium dodecyl sulfate-urea-gel electrophoresis, a molecular weight of about 96,000 was found for the phosphorylase II subunit. Molecular weight determination by nondenaturing gel electrophoresis at 5, 6, 7, and 8% acrylamide and by ultracentrifugation on sucrose density gradients suggested that the native enzyme is a dimer, as are other phosphorylases. © 1981. Fil:Sivak, M.N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Tandecarz, J.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00039861_v212_n2_p525_Sivak |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
isoenzyme phosphorylase article enzyme specificity enzymology heat isolation and purification kinetics metabolism molecular weight plant Heat Isoenzymes Kinetics Molecular Weight Phosphorylases Plants Substrate Specificity Support, Non-U.S. Gov't |
| spellingShingle |
isoenzyme phosphorylase article enzyme specificity enzymology heat isolation and purification kinetics metabolism molecular weight plant Heat Isoenzymes Kinetics Molecular Weight Phosphorylases Plants Substrate Specificity Support, Non-U.S. Gov't Sivak, M.N. Tandecarz, J.S. Cardini, C.E. Studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. I. Characterization and properties of the enzyme |
| topic_facet |
isoenzyme phosphorylase article enzyme specificity enzymology heat isolation and purification kinetics metabolism molecular weight plant Heat Isoenzymes Kinetics Molecular Weight Phosphorylases Plants Substrate Specificity Support, Non-U.S. Gov't |
| description |
Phosphorylase II from potato tuber has been subjected to several purification procedures in order to: (a) separate the unprimed activity from the primed one coexisting in the enzymatic preparation, and (b) eliminate endogenous glucan and/or glycoproteins. Large-scale analytical nondenaturing gel electrophoresis and affinity chromatography on concanavalin A-Sepharose 4B succeeded in removing endogenous glycoproteins without any effect on the unprimed activity. In addition, phosphorolysis or glucoamylase treatment of the enzymatic preparation did not abolish the phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. However, no separation between both activities, primed and unprimed, could be achieved either by the above-mentioned methods or by sucrose density gradient centrifugation. Based on sodium dodecyl sulfate-urea-gel electrophoresis, a molecular weight of about 96,000 was found for the phosphorylase II subunit. Molecular weight determination by nondenaturing gel electrophoresis at 5, 6, 7, and 8% acrylamide and by ultracentrifugation on sucrose density gradients suggested that the native enzyme is a dimer, as are other phosphorylases. © 1981. |
| format |
JOUR |
| author |
Sivak, M.N. Tandecarz, J.S. Cardini, C.E. |
| author_facet |
Sivak, M.N. Tandecarz, J.S. Cardini, C.E. |
| author_sort |
Sivak, M.N. |
| title |
Studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. I. Characterization and properties of the enzyme |
| title_short |
Studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. I. Characterization and properties of the enzyme |
| title_full |
Studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. I. Characterization and properties of the enzyme |
| title_fullStr |
Studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. I. Characterization and properties of the enzyme |
| title_full_unstemmed |
Studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. I. Characterization and properties of the enzyme |
| title_sort |
studies on potato tuber phosphorylase-catalyzed reaction in the absence of an exogenous acceptor. i. characterization and properties of the enzyme |
| url |
http://hdl.handle.net/20.500.12110/paper_00039861_v212_n2_p525_Sivak |
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