Zinc stoichiometry in Escherichia coli alkaline phosphatase. Studies by 31P NMR and ion-exchange chromatography
| Autor: | Jay L. Bock, Arthur Kowalsky |
|---|---|
| Rok vydání: | 1978 |
| Předmět: |
inorganic chemicals
Magnetic Resonance Spectroscopy Macromolecular Substances Ion chromatography chemistry.chemical_element Protein dimer Zinc medicine.disease_cause chemistry.chemical_compound Apoenzymes Escherichia coli medicine HEPES chemistry.chemical_classification Chromatography Chemistry General Medicine Hydrogen-Ion Concentration Alkaline Phosphatase Chromatography Ion Exchange Phosphate Enzyme biological sciences bacteria Alkaline phosphatase Protons Protein Binding Nuclear chemistry |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Enzymology. 526:135-146 |
| ISSN: | 0005-2744 |
| DOI: | 10.1016/0005-2744(78)90298-x |
| Popis: | 31P nuclear magnetic resonance spectra and enzymatic activities are compared for alkaline phosphatase (orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1) species with different zinc contents. The enzyme containing two Zn2+ per protein dimer exists in two forms; one, prepared by dialysis of native enzyme, has full enzymatic activity and a 31P magnetic resonance spectrum similar to but distinguishable from that of the native enzyme containing four or more Zn2+. The other form, prepared by restoring two Zn2+ to apoenzyme, has low enzymatic activity and a 31P magnetic resonance spectrum that indicates stoichiometric binding of phosphate, but otherwise altered properties. Reconstituted enzyme with four Zn2+ is similar to but distinguishable from native enzyme with four Zn2+. Chromatography on DEAE-cellulose can separate apoenzyme and enzyme containing two Zn2+ and suggests that the binding of a pair of Zn2+ is cooperative. |
| Databáze: | OpenAIRE |
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