人卵丘细胞中乙酰辅酶A：芳胺N - 乙酰转移酶的动力学

Kinetics of acetyl coenzyme A: arylamine N-acetyltransferase from human cumulus cells.

作者信息

Chang C C, Hsieh Y Y, Chung J G, Tsai H D, Tsai C H

机构信息

Department of Obstetrics and Gynecology, China Medical College Hospital, No. 2 Yuh-Der Road, Taichung, Taiwan.

出版信息

J Assist Reprod Genet. 2001 Sep;18(9):512-8. doi: 10.1023/a:1016605211332.

DOI:10.1023/a:1016605211332

PMID:11665667

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3455732/

Abstract

PURPOSE

N-acetyltransferase (NAT) activity is involved in the detoxification of exogenous amines. We aimed to evaluate the kinetics of acetyl coenzyme A (AcCoA): arylamine NAT for human cumulus cells.

METHODS

Thirty infertile women who were undergoing controlled ovarian hyperstimulation (COH) and oocyte retrievals were recruited. Human cumulus cells were obtained during oocyte retrievals. Using 2-aminofluorene (2-AF) and p-aminobenzoic acid (PABA) as substrates, NAT activity and Michaelis-Menten kinetics constants of all samples were determined by using high-pressure liquid chromatography.

RESULTS

There were 6 rapid, 10 intermediate, and 14 slow acetylators. 2-AF-NAT and PABA-NAT activities were 0.97 +/- 0.74 and 0.89 +/- 0.77 nmol/min/mg protein, respectively. Km/Vmax of rapid and slow acetylators for 2-AF were (161 +/- 55)/(15.6 +/- 2.9) and (27.8 +/- 11.4)/(2.6 +/- 0.9), respectively. Km/Vmax of rapid and slow acetylators for PABA were (104 +/- 36)/(13.2 +/- 2.8) versus (20.0 +/- 10)/(2.0 +/- 0.7), respectively. Compared to slow acetylators, the rapid acetylators exhibited higher Km/Vmax values for 2-AF (5.8-/6-fold) and PABA (6-/6.6-fold), respectively.

CONCLUSION

Human cumulus could acetylate arylamine carcinogen (2-AF) and noncarcinogen drug (PABA). Higher percentage of rapid acetylators established in the cumulus during COH. It provides a model for monitoring the effects of pollution or carcinogenesis upon the oocyte during COH and oocyte retrievals.

摘要

目的

N - 乙酰转移酶（NAT）活性参与外源性胺的解毒过程。我们旨在评估人卵丘细胞中乙酰辅酶A（AcCoA）：芳胺NAT的动力学。

方法

招募了30名正在接受控制性卵巢过度刺激（COH）和卵母细胞采集的不孕女性。在卵母细胞采集过程中获取人卵丘细胞。以2 - 氨基芴（2 - AF）和对氨基苯甲酸（PABA）为底物，使用高压液相色谱法测定所有样本的NAT活性和米氏动力学常数。

结果

有6名快速乙酰化者、10名中间型乙酰化者和14名缓慢乙酰化者。2 - AF - NAT和PABA - NAT活性分别为0.97±0.74和0.89±0.77 nmol/分钟/毫克蛋白。快速和缓慢乙酰化者对2 - AF的Km/Vmax分别为(161±55)/(15.6±2.9)和(27.8±11.4)/(2.6±0.9)。快速和缓慢乙酰化者对PABA的Km/Vmax分别为(104±36)/(13.2±2.8)和(20.0±10)/(2.0±0.7)。与缓慢乙酰化者相比，快速乙酰化者对2 - AF和PABA的Km/Vmax值分别更高（5.8 - /6倍）和（6 - /6.6倍）。