AI Article Synopsis
- NADH plays a crucial role as a cofactor in various metabolic reactions, particularly influencing glycolysis through its redox state in the cytosol.
- Traditional methods for measuring NADH involve chemical tests or autofluorescence imaging, which are unable to target NADH levels specifically in the cytosol of live cells.
- The development of Peredox, a genetically encoded fluorescent biosensor, allows for more precise measurement of the cytosolic NADH-NAD(+) redox state in individual live cells, enhancing imaging techniques and providing important technical insights.
Article Abstract
NADH is an essential redox cofactor in numerous metabolic reactions, and the cytosolic NADH-NAD(+) redox state is a key parameter in glycolysis. Conventional NADH measurements rely on chemical determination or autofluorescence imaging, which cannot assess NADH specifically in the cytosol of individual live cells. By combining a bacterial NADH-binding protein and a fluorescent protein variant, we have created a genetically encoded fluorescent biosensor of the cytosolic NADH-NAD(+) redox state, named Peredox (Hung et al., Cell Metab 14:545-554, 2011). Here, we elaborate on imaging methods and technical considerations of using Peredox to measure cytosolic NADH:NAD(+) ratios in individual live cells.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4330558 | PMC |
| http://dx.doi.org/10.1007/978-1-62703-622-1_7 | DOI Listing |
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