Skip to main content
SpringerLink
Log in

A colorimetric method for the assay of lipoyl dehydrogenase

  • Published:
Microchimica Acta Aims and scope Submit manuscript

Summary

A colorimetric method for the direct assay of lipoyl dehydrogenase is described. Enzyme reaction is stopped by ethanol precipitation. This is followed by the displacement of 1,3-bis(2′-pyridyl)-1,2-diaza-prop-2-ene (PAPHY) from a Pd(II)-PAPHY complex by reduced lipoic acid. This simple quantitative method is suitable for assay of lipoyl dehydrogenase in crude tissue extracts and in cell suspensions.

Zusammenfassung

Eine kolorimetrische Methode zur direkten Bestimmung von Liponsäuredehydrogenase wurde beschrieben. Die enzymatische Reaktion wird durch Alkoholfällung unterbrochen. Danach verdrängt man mit Hilfe der reduzierten Liponsäure das 1,3-Bis-(2′-pyridyl)-1,2-diazapropen-2 (PAPHY) aus seiner Pd(II)-Komplexverbindung. Für die Bestimmung der Liponsäuredehydrogenase in Geweberohextrakten oder Zellsuspensionen eignet sich dieses einfache Verfahren gut.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

EDTA:

ethylenediaminotetraacetic acid

Lip S2 :

DL-6, 8-thioctic aicd

Lip(S)2 :

DL-6, 8-thioctic acid, reduced form

NAD:

β-Nicotinamide adenine dinucleotide

NADH:

β-Nicotinamide adenine dinucleotide, reduced form

References

  1. A. M. Seet and K. T. Lee, Mikrochim. Acta [Wien]1974, 235.

  2. V. Massey, Biochim. Biophys. Acta37, 314 (1960).

    Google Scholar 

  3. F. B. Sträub, Biochem. J.33, 787 (1939).

    Google Scholar 

  4. C. J. Lusty and T. P. Singer, J. Biol. Chem.239, 3733 (1964).

    Google Scholar 

  5. T. Hayakawa, Y. Sakurai, T. Aikawa, Y. Fukuyoshi, and M. Koike, in K. Yagi “Flavins and Flavoproteins” Tokyo: Univ. of Tokyo Press. 1968. p. 99.

    Google Scholar 

  6. J. Visser, H. Voetberg, and C. Veeger, in H. Sund: “Pyridine Nucleotide Dependent Dehydrogenases”. Berlin: Springer-Verlag. 1969. p. 359.

    Google Scholar 

  7. V. Massey, in W. A. Wood (ed.) “Methods in Enzymology” Vol. IX, New York: Academic Press. 1966. p. 272.

    Google Scholar 

  8. T. C. Tsao and K. Bailey, Biochim. Biophys. Acta11, 102 (1953).

    Google Scholar 

  9. V. Massey, in P. D. Boyer, H. Lardy, and K. Myrback, “The Enzymes” Vol. 7, New York: Academic Press. 1963. p. 275.

    Google Scholar 

Download references

Author information

Author notes

  1. Kum -Tatt Lee

    Present address: Singapore Institute of Standards and Industrial Research, 179 River Valley Road, Singapore 6, Republic of Singapore

Authors and Affiliations

  1. Government Department of Pathology, Clinical Biochemistry Laboratories, Outram Road, Singapore 3, Republic of Singapore

    Ai -Mee Seet & Kum -Tatt Lee

Authors
  1. Ai -Mee Seet
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kum -Tatt Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Seet, A.M., Lee, K.T. A colorimetric method for the assay of lipoyl dehydrogenase. Mikrochim Acta 63, 577–583 (1975). https://doi.org/10.1007/BF01217319

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01217319

Keywords

Search

Navigation