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成年黑腹果蝇谷胱甘肽S-转移酶蛋白质组的研究:对化学刺激的反应性。

Studies on the glutathione S-transferase proteome of adult Drosophila melanogaster: responsiveness to chemical challenge.

作者信息

Alias Zazali, Clark Alan G

机构信息

School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand.

出版信息

Proteomics. 2007 Oct;7(19):3618-28. doi: 10.1002/pmic.200700070.

Abstract

GSTs from adult Drosophila melanogaster have been partially purified using three different affinity chromatography media and separated by 2-DE. Nine GSTs have been identified by MALDI-TOF MS. In the absence of special treatments, eight GSTs could be positively identified. These were DmGSTs D1 (the dominant Delta isoform which was present in five protein zones of differing pI) and D3 (and possibly also D5); the Epsilon-class GSTs E3, 6, 7 and 9 and a previously uncharacterised, probable member of the class, CG16936. The Sigma-class DmGSTS1 was prominent. DmGSTD2 was detected only after pretreatment of the flies with Phenobarbital (PhB). Treatment with Paraquat (PQ) led to an increase in the total GST activity, as measured with the substrates 1-chloro-2,4-dinitrobenzene (CDNB) and 3,4-dichloro-nitrobenzene (DCNB) and an increase in the relative amounts of the D1, D3, E6 and E7 isoforms. PhB treatment led to increases in the relative amounts of the D1, D2, E3, E6, E7 and E9 isoforms detected with a possible depression in the relative amount of GSTS1. CG16936 was unaffected by either pretreatment.

摘要

利用三种不同的亲和层析介质对成年黑腹果蝇的谷胱甘肽S-转移酶(GSTs)进行了部分纯化,并通过双向电泳(2-DE)进行分离。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定出了9种GSTs。在没有特殊处理的情况下,可以明确鉴定出8种GSTs。它们分别是DmGSTs D1(占主导地位的Delta亚型,存在于5个不同等电点的蛋白区域)和D3(可能还有D5);Epsilon类GSTs E3、6、7和9,以及该类中一个此前未鉴定的可能成员CG16936。Sigma类的DmGSTS1很突出。仅在对果蝇用苯巴比妥(PhB)预处理后才检测到DmGSTD2。用百草枯(PQ)处理导致以1-氯-2,4-二硝基苯(CDNB)和3,4-二氯硝基苯(DCNB)为底物测定的总GST活性增加,以及D1、D3、E6和E7亚型的相对含量增加。PhB处理导致检测到的D1、D2、E3、E6、E7和E9亚型的相对含量增加,而GSTS1的相对含量可能降低。CG16936不受任何一种预处理的影响。

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