Abstract |
3-Phosphoglycerate kinase (3-PGK) has been purified to apparent homogeneity from Ehrlich ascites carcinoma (EAC) cells by (NH4)2SO4 precipitation, gel filtration and ion-exchange chromatography. The enzyme has been partially characterized and compared with the characteristics of this enzyme of other normal and malignant cells. The EAC cell 3-PGK is composed of a single subunit of 47 kDa. It has a broad pH optimum (pH 6.0-7.5) for its enzymatic activity. The apparent Km values of 3-phosphoglycerate (3-PGA) and ATP for 3-PGK have been found out to be 0.25 mM and 0.1 mM respectively. Similar to 3-PGK of other cells, the EAC enzyme requires either Mg2+ or Mn2+ for full activity; the optimum concentrations of Mg2+ and Mn2+ are 0.8 mM and 0.5 mM respectively. When ATP and 3-PGA act as substrates, ADP, the reaction product of 3-PGK-catalyzed reaction has been found to inhibit this enzyme. Kinetic studies were made on the inhibition of ADP in presence of the substrates ATP and 3-PGA. Attempts to hybridize 3-PGK and glyceraldehyde-3-phosphate dehydrogenase of EAC cells by NAD or glutaraldehyde were unsuccessful.
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Authors | Kasturi Mukherjee, Swapna Ghosh, Manju Ray, Subhankar Ray |
Journal | Indian journal of biochemistry & biophysics
(Indian J Biochem Biophys)
Vol. 39
Issue 5
Pg. 332-41
(Oct 2002)
ISSN: 0301-1208 [Print] India |
PMID | 22905379
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Manganese
- Mercaptoethanol
- Adenosine Diphosphate
- Adenosine Triphosphate
- Edetic Acid
- Phosphoglycerate Kinase
- Magnesium
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Topics |
- Adenosine Diphosphate
(chemistry)
- Adenosine Triphosphate
(metabolism)
- Animals
- Carcinoma, Ehrlich Tumor
(metabolism)
- Chromatography, Gel
- Chromatography, Ion Exchange
(methods)
- Edetic Acid
(chemistry)
- Gene Expression Regulation, Enzymologic
- Gene Expression Regulation, Neoplastic
- Hydrogen-Ion Concentration
- Kinetics
- Magnesium
(chemistry)
- Manganese
(chemistry)
- Mercaptoethanol
(pharmacology)
- Mice
- Neoplasm Transplantation
- Nucleic Acid Hybridization
- Phosphoglycerate Kinase
(chemistry, isolation & purification)
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