Microsomal acetoxy drug: protein transacetylase of placenta: part 1. Characterization of DAMC: GST transacetylase.
We have earlier established in tissues of several mammalian animal species the existence of a novel membrane bound enzyme termed 7,8-diacetoxy-4-methylcoumarin (DAMC): protein transacetylase (TAase) that possibly transfers acetyl groups from the model acetoxy drug (DAMC) to certain enzyme protein viz. glutathione S-transferase (GST), cytochrome P-450 and NADPH cytochrome C reductase leading to the drastic modulation of their catalytic activities. We have in this report extended the studies to human tissue and characterized TAase from placenta. For this purpose placental microsomes were preincubated with DAMC along with the receptor protein (cytosolic GST) followed by the addition of the substrates of GST in order to quantify the catalytic activity of GST, the extent of inhibition of GST served as a measure of TAase. Placental TAase was also found to irreversibly activate NADPH cytochrome C reductase by DAMC. Placental enzyme activated the reductase even at very low concentration of DAMC. Iodoacetamide nearly abolished the placental TAase suggesting the presence of active thiol group in the enzyme and the TAase demonstrated hyperbolic kinetics. Kinetic constants obtained by varying the concentrations of either of the substrates DAMC or cytosolic GST characterized TAase catalysed reaction as the bimolecular reaction. Further studies are in progress to delineate the physiological importance of TAase in placenta.
Kumari
R
R
Department of Biochemistry, V. P. Chest Institute, University of Delhi, Delhi, 110 007 India.
Kohli
E
E
Gupta
G
G
Raj
H G
HG
Parmar
V S
VS
Pal
G
G
Tyagi
Y K
YK
Jain
S C
SC
Olsen
C E
CE
D000123:Acetyltransferases / Q000378:metabolism*; D000328:Adult; D000818:Animals; D003374:Coumarins / Q000378:metabolism*; D005260:Female; D005978:Glutathione / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D051379:Mice; D008861:Microsomes / Q000201:enzymology; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity
C114369:7,8-diacetoxy-4-methylcoumarin; D003374:Coumarins; D009251:NADPH-Ferrihemoprotein Reductase; D000123:Acetyltransferases; D005982:Glutathione Transferase; D005978:Glutathione