Evidence for the Essential Arginine and Histidine Residues in Catalytic Activity of Glucose 6-Phosphate Dehydrogenase from Streptomyces aureofaciens
Glucose 6-phosphate dehydrogenase (G6PD) was purified from Streptomyces aureofaciens and inactivated with butanedione and diethylpyrocarbonate. Incubation of the enzyme with butanedione resulted in a rapid activity loss (80%) within 5 min, followed by a slow phase using a molar ratio to enzyme conce...
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| Language: | English |
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University of Tehran
2005-03-01
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| Series: | Journal of Sciences, Islamic Republic of Iran |
| Online Access: | https://jsciences.ut.ac.ir/article_31621_9113301d6db6f4521430630c7ae3c7ec.pdf |
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| description | Glucose 6-phosphate dehydrogenase (G6PD) was purified from Streptomyces aureofaciens and inactivated with butanedione and diethylpyrocarbonate. Incubation of the enzyme with butanedione resulted in a rapid activity loss (80%) within 5 min, followed by a slow phase using a molar ratio to enzyme concentration of 100. Fluorescence studies showed a conformational change in the butanedione-modified enzyme. NAD+, NADP+ and glucose 6-phosphate protected the enzyme against inactivation. Diethylpyrocarbonate (2 mM) completely inactivated the enzyme after 2 min. Stoichiometry of the inactivation showed 2 moles of histidine residues per mole of enzyme with complete activity loss. Maximum emission spectrum of the enzyme decreased (23%) upon modification and the presence of NAD+ or NADP+ further decreased the fluorescence by 27% and 10.5%, respectively. The data suggest that essential arginine and histidine residues may be involved in the catalytic activity of Streptomyces aureofaciens G6PD |
| format | Article |
| id | doaj-art-3a32ce83d2a64d84aac2265dc9c4fb78 |
| institution | OA Journals |
| issn | 1016-1104 2345-6914 |
| language | English |
| publishDate | 2005-03-01 |
| publisher | University of Tehran |
| record_format | Article |
| series | Journal of Sciences, Islamic Republic of Iran |
| spelling | doaj-art-3a32ce83d2a64d84aac2265dc9c4fb782025-08-20T01:53:33ZengUniversity of TehranJournal of Sciences, Islamic Republic of Iran1016-11042345-69142005-03-0116131621Evidence for the Essential Arginine and Histidine Residues in Catalytic Activity of Glucose 6-Phosphate Dehydrogenase from Streptomyces aureofaciensGlucose 6-phosphate dehydrogenase (G6PD) was purified from Streptomyces aureofaciens and inactivated with butanedione and diethylpyrocarbonate. Incubation of the enzyme with butanedione resulted in a rapid activity loss (80%) within 5 min, followed by a slow phase using a molar ratio to enzyme concentration of 100. Fluorescence studies showed a conformational change in the butanedione-modified enzyme. NAD+, NADP+ and glucose 6-phosphate protected the enzyme against inactivation. Diethylpyrocarbonate (2 mM) completely inactivated the enzyme after 2 min. Stoichiometry of the inactivation showed 2 moles of histidine residues per mole of enzyme with complete activity loss. Maximum emission spectrum of the enzyme decreased (23%) upon modification and the presence of NAD+ or NADP+ further decreased the fluorescence by 27% and 10.5%, respectively. The data suggest that essential arginine and histidine residues may be involved in the catalytic activity of Streptomyces aureofaciens G6PDhttps://jsciences.ut.ac.ir/article_31621_9113301d6db6f4521430630c7ae3c7ec.pdf |
| spellingShingle | Evidence for the Essential Arginine and Histidine Residues in Catalytic Activity of Glucose 6-Phosphate Dehydrogenase from Streptomyces aureofaciens Journal of Sciences, Islamic Republic of Iran |
| title | Evidence for the Essential Arginine and Histidine Residues in Catalytic Activity of Glucose 6-Phosphate Dehydrogenase from Streptomyces aureofaciens |
| title_full | Evidence for the Essential Arginine and Histidine Residues in Catalytic Activity of Glucose 6-Phosphate Dehydrogenase from Streptomyces aureofaciens |
| title_fullStr | Evidence for the Essential Arginine and Histidine Residues in Catalytic Activity of Glucose 6-Phosphate Dehydrogenase from Streptomyces aureofaciens |
| title_full_unstemmed | Evidence for the Essential Arginine and Histidine Residues in Catalytic Activity of Glucose 6-Phosphate Dehydrogenase from Streptomyces aureofaciens |
| title_short | Evidence for the Essential Arginine and Histidine Residues in Catalytic Activity of Glucose 6-Phosphate Dehydrogenase from Streptomyces aureofaciens |
| title_sort | evidence for the essential arginine and histidine residues in catalytic activity of glucose 6 phosphate dehydrogenase from streptomyces aureofaciens |
| url | https://jsciences.ut.ac.ir/article_31621_9113301d6db6f4521430630c7ae3c7ec.pdf |