Studies on glutathione transferase from grasshopper (Zonocerus variegatus)

被引:11
作者
Adewale, IO [1 ]
Afolayan, A [1 ]
机构
[1] Obafemi Awolowo Univ, Dept Biochem, Ife, Nigeria
关键词
grasshopper; glutathione transferase; polyphagous; substrates; inhibition; NAD; NADH; kinetics;
D O I
10.1016/j.pestbp.2005.10.004
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Glutathione transferase (GST) was purified from the hindgut of grasshopper (Zonocerus variegatus) a polyphagous insect. The purified enzyme had a native molecular weight of 40 kDa and a subunit molecular weight of 19 kDa. The purified enzyme could conjugate glutathione (GSH) with 1-chloro-2,4-dinitrobenzene (CDNB), paranitrobenzylchloride, paranitrophenylacetate, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBDCl), and 1,2-dichloro-4-nitrobenzene (DCNB) with specific activities of 3.3 +/- 0.3, 0.49 +/- 0.10, 0.10 +/- 0.002, 1.2 +/- 0.2, and 1.7 +/- 0.4 mu mol/min/mg protein, respectively. CDNB appears to be the best substrate with a specificity constant, k(cat)/K-m, of 1.8 +/- 0.1 x 10(-4) M-1 S-1. The kinetic mechanism of Z. variegatus GST (zvGST) in the conjugation of GSH with some electrophilic substrates appears complex. Conjugation of GSH with DCNB was inhibited by high DCNB concentration, while with NBDCl, as the electrophilic substrates, different values of K-m were obtained at high and low concentrations of the substrates. Cibacron blue, hematin, S-hexylglutathione, and oxidized glutathione inhibited the enzyme with 150 values of 0.057 +/- 0.004, 0.80 +/- 0.2, 33 +/- 2 mu M, and 5.2 +/- 0.3 mM, respectively. The nature of inhibition by each of these inhibitors is either competitive or non-competitive at varying GSH or CDNB as substrates. NADH and NAD+ inhibited the enzyme with an 150 value of 0.4 +/- 0.01 and 11 +/- 1 mM, respectively. NADH at a concentration of 0.54 mM completely abolished the activity. As part of its adaptation, the flexible kinetic pathway of detoxication by zvGST may assist the organism in coping with various xenobiotics encountered in its preferred food plants. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:52 / 59
页数:8
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