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Organelle sensor: amperometric determination of nadh by immobilized mitochondrial electron transport particles

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Abstract

A new sensor for NADH was developed by making use of an immobilized subcellular organelle. Mitochondria was used as a model system for assembling an organelle sensor. Mitochondrial electron transport particles (ETP) were prepared from beef heart muscle and entrapped in the membrane formed of agar gel. The membrane-bound ETP was found capable of NADH oxidation:

$$NADH + \tfrac{1}{2}O_2 + H^ + \xrightarrow{{ETP}}NAD^ - + H_2 O$$

The membrane was tightly attached to the surface of an oxygen electrode capable of amperometric detection of O2. The sensor responded to NADH in solution with a resulting electric output. The response was enhanced by the addition of 2,4-dinitrophenol (DNP). NADH was determined in the concentration range 1–300 µM. NADH was alternatively determined for 2 weeks without replacing the ETP-bound membrane.

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Authors and Affiliations

  1. Research Laboratory of Resources Utilization, Tokyo Institute of Technology Nagatsuta, Modiri-ku, Yokohama 227, Japan

    Masuo Aizawa, Mitsuo Wada & Shuichi Suzuki

Authors
  1. Masuo Aizawa
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  2. Mitsuo Wada
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  3. Shuichi Suzuki
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Aizawa, M., Wada, M. & Suzuki, S. Organelle sensor: amperometric determination of nadh by immobilized mitochondrial electron transport particles. Journal of Solid-Phase Biochemistry 5, 35–44 (1980). https://doi.org/10.1007/BF02991901

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  • DOI: https://doi.org/10.1007/BF02991901

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