In vitro 6xHis-oMLE activity assays for ME, LDH and MDH function, using the oMLE storage buffer as assay buffer.
2021
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Zugriff:
(A) ME activity was observed for 6xHis-oMLE in a ME activity assay containing 10 mM malate, 1 mM NAD + and 1 mM. Omission of MnCl 2 reduced activity. (B) MDH activity was observed for 6xHis-oMLE in an assay containing 5 mM malate and 0.5 mM NADH. Activity was greatly increased by the addition of 1 mM MnCl 2 . (D) LDH activity was exhibited by 6xHis-oMLE in an LDH assay containing 5 mM pyruvate and 0.5 mM NADH. Activity was unchanged by the addition of 1 mM MnCl 2 . All assays used 28.5 μg 6xHis-oMLE per well or the equivalent volume of 6xHis-mock purification negative control and were performed at 45°C in 100 mM HEPES/0.1 mMnCl 2 pH 6.0 buffer. Activity was measured using increase or decrease of light absorbance at 340 nm, correlating to NADH oxidation and NAD + reduction respectively during enzyme activity. Error bars represent standard deviation around the mean from at least three independent experiments. (TIF)
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In vitro 6xHis-oMLE activity assays for ME, LDH and MDH function, using the oMLE storage buffer as assay buffer.
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Autor/in / Beteiligte Person: | Janine Hofmann (11273418) ; Mebratu A. Bitew (11273421) ; Miku Kuba (11273424) ; David P. De Souza (2793088) ; Hayley J. Newton (8692020) ; Fiona M. Sansom (8893082) |
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Veröffentlichung: | 2021 |
Medientyp: | Bild |
DOI: | 10.1371/journal.pone.0255925.s004 |
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