Skip to main content
SpringerLink
Log in

A rapid measurement of AΔ5-3β-hydroxysteroid dehydrogenase activity coupled with Δ4−Δ5 isomerase by a high pressure liquid chromatography

  • Short Communication
  • Published:
Journal of Endocrinological Investigation Aims and scope Submit manuscript

Abstract

By a high pressure liquid chromatography, progesterone (λmax = 240 nm, ε240 = 17,000) was separable from the other Δ4-3-oxosteroids. Relationship between the amount of progesterone and the area of the peak corresponding to progesterone was established as linear at least between 0–2.0 g. Progesterone is formed from pregnenolone by Δ5-3β-hydroxysteroid dehydrogenase coupled with Δ4−Δ5 isomerase. Because pregnenolone is devoid of absorption at 240 nm, the enzyme activity was measurable by the high pressure liquid chromatography from the amount of progesterone produced from pregnenolone in vitro in the presence of nicotinamide adenine dinucleotide. As an example of its application, the dehydrogenase activities in the ovaries of the immature rats treated with gonadotropins were measured by the above method. The estimates obtained by this method were agreeable with those by thin layer chromatography, employing radioactive pregnenolone as the substrate.

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

References

  1. Lamber P.W., Syverson B.J., Arnaud C.D., Speisberg T.C. Isolation and quantitation of endogenous vitamin D and its physiologically important metabolites in human plasma by high pressure liquid chromatography. J. Steroid Biochem. 8: 929, 1977.

    Article  Google Scholar 

  2. Suzuki K., Kawakura K., Tamaoki B. Effect of pregnant mare’s serum gonadotropin on the activities of Δ4−5β-reductase, aromatase, and other enzymes in the ovaries of immature rats. Endocrinology 102: 1595, 1978.

    Article  CAS  PubMed  Google Scholar 

  3. Suzuki K., Tamaoki B. Enzymological studies of rat luteinized ovaries in relation to acute reduction of aromatizable androgen formation and stimulated production of progestins. Endocrinology 104: 1317, 1979.

    Article  CAS  PubMed  Google Scholar 

  4. Bryson M.J., Sweat M.L. Metabolism of progesterone in human myometrium. Endocrinology 84: 1071, 1969.

    Article  CAS  PubMed  Google Scholar 

  5. Taylor R.B., Kendle K.E. Use of HPLC to determine the effect of 17β-hydroxy-7-methyl-androst-5-en-3-one (RMI 12,936) on production of progesterone by rat ovarian homogenate. Acta Endocrinol. (Kbh.) 88: 157, 1978.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. National Institute of Radiological Sciences, 9-1 Anagawa-4-chome, Chiba-shi 260, Japan

    Keiko Suzuki & B. Tamaoki

  2. Kyoritsu College of Pharmacy, Shiba Park, Minato-ku, Tokyo 105, Japan

    A. Kadowaki

Authors
  1. Keiko Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Kadowaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. B. Tamaoki
    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

Suzuki, K., Kadowaki, A. & Tamaoki, B. A rapid measurement of AΔ5-3β-hydroxysteroid dehydrogenase activity coupled with Δ4−Δ5 isomerase by a high pressure liquid chromatography. J Endocrinol Invest 3, 441–444 (1980). https://doi.org/10.1007/BF03349387

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03349387

Key-words

Search

Navigation