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J. Biochem, 1984, Vol. 95, No. 6 1733-1739
© 1984 Japanese Biochemical Society


research-article

Hydroxylation of Prostaglandin A1 by the Microsomes of Rabbit Intestinal Mucosa1

Emi KUSUNOSE*, Masatoshi KAKU*, Kosuke ICHIHARA**, Satoru YAMAMOTO*, Ikuya YANO*** and Masamichi KUSUNOSE*

*Toneyama Institute for Tuberculosis Research, Osaka City University Medical School Toyonaka, Osaka 560
**Department of Biochemistry, Kawasaki Medical School Kurashiki, Okayama 701-01
***Department of Bacteriology, Niigata University Medical School Niigata, Niigata 951

Microsomes from rabbit small intestine mucosa were found to catalyze the hydroxylation of PGA1 in the presence of NADPH. The major product was identified as 20-hydroxy PGAI by using high performance liquid chromatography and gas chromatography-mass spectrometry, and the minor product was assumed to be 19-hydroxy PGA1. The ratio of the former product to the latter was about 24.1. The specific PGA1 {omega}-hydroxylase activity of small intestine microsomes was comparable to that of liver microsomes, and was significantly higher than those of microsomes from other tissues such as kidney cortex and lung. Microsomes from rabbit colon mucosa also catalyzed the hydroxylation of PGA1 in the presence of NADPH, with the ratio of {omega}- to ({omega}-1)-hydroxy PGA1 formed being 33.0. The PGA1 hydroxylase activities of the microsomes from both small intestine and colon were inhibited markedly by carbon monoxide, indicating the participation of cytochrome P-450. A cytochrome P-450 was solubilized from small intestine microsomes, and purified to a specific content of 10.5 nmol of cytochrome P-450/mg of protein. This cytochrome hydroxylated PGA1 at the {omega}-position with a turnover rate of 38.2 nmol/min/nmol of cytochrome P-450 in the reconstituted system containing cytochrome P-450, NADPH-cytochrome P-450 reductase, cytochrome b5 and phosphatidyicholine. It is suggested that this cytochrome P-450 is specialized for the {omega}-hydroxylation of PGA1 in small intestine microsomes.

1Communications should be addressed to: M. Kusunose, Toneyama Institute for Tuberculosis Research, Toneyama 5-1-1, Toyonaka, Osaka 560.


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