Solubilisation and reconstitution of acylcoenzyme A:estradiol-17β acyltransferase

https://doi.org/10.1016/S0006-291X(87)80004-9Get rights and content

Summary

Acylcoenzyme A:estradiol-17β acyltransferase in microsomes of bovine placenta cotyledons was strongly membrane bound. The enzyme was solubilised from microsomes by sodium cholate and was reconstituted into phospholipid vesicles. The apparent Km for estradiol-17β was 11 μM which was close to the value of 8 μM previously found with the membrane-bound enzyme. Testosterone was also a substrate for the reconstituted enzyme (apparent Km 62 μM) and was a competitive inhibitor (Ki 74 μM) of the acylation of estradiol-17β. Although various long-chained fatty acyl CoAs acted as acyl donors, these proved to have widely differing apparent Km values with palmitoleoyl CoA having the highest affinity (Km 24 μM) and arachidonoyl CoA the lowest affinity (Km 330 μM).

References (13)

  • Mellon-NussbaumS.H. et al.

    J. Biol. Chem.

    (1982)
  • LarnerJ.M. et al.

    J. Steroid Biochem.

    (1985)
  • Abul-HajjY. et al.

    Steroids

    (1983)
  • AdamsJ.B. et al.

    J. Steroid Biochem.

    (1986)
  • BradfordM.M.

    Analytical Biochem.

    (1976)
  • SpectorA.A. et al.

    Prog. Lipid Res.

    (1979)
There are more references available in the full text version of this article.

Cited by (0)

View full text