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J. Biochem, 1988, Vol. 104, No. 1 35-39
© 1988 Japanese Biochemical Society

research-article

NMR-Invisible ATP in Rat Heart and Its Change in Ischemia1

Hiroshi Takami*, Eisuke Furuya**, Kunio Tagawa**,2, Yoshiteru Seo***, Masataka Murakami***, Hiroshi Watari***, Hikaru Matsuda*, Hajime Hirose* and Yasunaru Kawashima*

*First Department of Surgery, Osaka University Medical School Kita-ku, Osaka, Osaka 530
**Department of Physiological Chemistry, Osaka University Medical School Kita-ku, Osaka, Osaka 530
***Department of Molecular Physiology, National Institute for Physiological Sciences Okazaki, Aichi 444

2 To whom correspondence should be addressed

The subcellular compartmentalization of adenosine 5'-triphosphate (ATP) in isolated perfused rat heart and its relation to energy depletion in ischemia were examined by 31P nuclear magnetic resonance (31P-NMR) spectroscopy and chemical analyses. The signal intensities of the ß-phosphate of ATP and creatine phosphate in the 31P-NMR were standardized by the intracellular volume ratio measured with 23Na-NMR to determine the actual content of each. During aerobic perfusion the ATP content determined by NMR (13.7 ± 2.2 µmol/g dry weight) was significantly lower than that found by chemical analysis (22.4±0.7 µmol/g dry weight), while the creatine phosphate contents determined by the two methods were the same. During ischemia at 33°C the signal of the ß-phosphate of ATP in the 31PNR spectrum decreased progressively, disappearing completely after 16 min. But at this time 5.7± 1.7 µmo1/g dry weight of myocardial ATP was still detected by chemical analysis. These results indicated that there were two different compartments of intracellular ATP in the heart, only one of which is detectable by 31P-NMR spectroscopy, and that during ischeniia the ATP that is detectable, which seems to be the free ATP in the cytosol, decreased more rapidly than the ATP in the other compartment.

1 This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.


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