Enzymatic androgen assay: some properties of human placental microsomal aromatase.
The androgen content of biological fluids can be determined after their conversion into estrogens using human placental microsomal aromatase (HPMA). The purpose of this paper is to report some physico-chemical properties of HPMA. Using an accurate, specific and sensitive assay for HPMA, Km values for dehydroepiandrosterone (DHEA), androstenedione and testosterone were found to increase with increasing amount of the detergent (Triton X-100) added. Analysis at substrate concentrations 5-10 times above and below the Km values did not indicate any anomalous kinetic behaviour. Triton X-100, used for enzyme solubilization, significantly decreased the rate of aromatization of the three substrates by increasing their Km values. This effect was more important for testosterone than for androstenedione or DHEA. Using a new protocol for the determination of aromatase activity, kinetic properties of aromatase before and after solubilization are described.
Chikhaoui
Y
Y
Chitour
F
F
Boussioux
A M
AM
Nicolas
J C
JC
Descomps
B
B
Crastes de Paulet
A
A
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000735:Androstenedione / Q000378:metabolism; D001141:Aromatase / Q000378:metabolism*; D003687:Dehydroepiandrosterone / Q000378:metabolism; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D009249:NADP / Q000378:metabolism; D017830:Octoxynol; D010088:Oxidoreductases / Q000378:metabolism*; D010920:Placenta / Q000201:enzymology*; D011092:Polyethylene Glycols; D011247:Pregnancy; D012995:Solubility; D013739:Testosterone / Q000378:metabolism
D011092:Polyethylene Glycols; D013739:Testosterone; D000735:Androstenedione; D003687:Dehydroepiandrosterone; D009249:NADP; D017830:Octoxynol; D010088:Oxidoreductases; D015096:3-Hydroxysteroid Dehydrogenases; D001141:Aromatase