Conformational dependence of intracellular NADH on metabolic state revealed by associated fluorescence anisotropy

Harshad D. Vishwasrao, Ahmed A. Heikal, Karl A. Kasischke, Watt W. Webb

Research output: Contribution to journalArticlepeer-review

233 Scopus citations


Global analysis of fluorescence and associated anisotropy decays of intrinsic tissue fluorescence offers a sensitive and non-invasive probe of the metabolically critical free/enzyme-bound states of intracellular NADH in neural tissue. Using this technique, we demonstrate that the response of NADH to the metabolic transition from normoxia to hypoxia is more complex than a simple increase in NADH concentration. The concentration of free NADH, and that of an enzyme bound form with a relatively low lifetime, increases preferentially over that of other enzyme bound NADH species. Concomitantly, the intracellular viscosity is reduced, likely due to the osmotic swelling of mitochondria. These conformation and environmental changes effectively decrease the tissue fluorescence average lifetime, causing the usual total fluorescence increase measurements to significantly underestimate the calculated concentration increase. This new discrimination of changes in NADH concentration, conformation, and environment provides the foundation for quantitative functional imaging of neural energy metabolism.

Original languageEnglish (US)
Pages (from-to)25119-25126
Number of pages8
JournalJournal of Biological Chemistry
Issue number26
StatePublished - Jul 1 2005

Bibliographical note

Copyright 2008 Elsevier B.V., All rights reserved.

Fingerprint Dive into the research topics of 'Conformational dependence of intracellular NADH on metabolic state revealed by associated fluorescence anisotropy'. Together they form a unique fingerprint.

Cite this