期刊
ENVIRONMENTAL POLLUTION
卷 157, 期 8-9, 页码 2459-2469出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.envpol.2009.03.015
关键词
Glutathione transferase; Lumbricus rubellus; Earthworm; 2-DE
资金
- NERC UK (Environmental Genomics [NER/T/S/2002/00021]
- Natural Environment Research Council [CEH010021, ceh010023, NER/T/S/2002/00021] Funding Source: researchfish
The earthworm Lumbricus rubellus (Hoffmeister, 1843) is a terrestrial pollution sentinel. Enzyme activity and transcription of phase II detoxification superfamily glutathione transferases (GST) is known to respond in earthworms after soil toxin exposure, suggesting GST as a candidate molecular-based pollution biomarker. This study combined sub-proteomics, bioinformatics and biochemical assay to characterise the L. rubellus GST complement as pre-requisite to initialise assessment of the applicability of GST as a biomarker. L rubellus possesses a range of GSTs related to known classes, with evidence of tissue-specific synthesis. Two affinity-purified GSTs dominating GST protein synthesis (Sigma and Pi class) were cloned, expressed and characterised for enzyme activity with various substrates. Electrospray ionisation mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) following SDS-PAGE were superior in retaining subunit stability relative to two-dimensional gel electrophoresis (2-DE). This study provides greater understanding of Phase II detoxification GST superfamily status of an important environmental pollution sentinel organism. (c) 2009 Elsevier Ltd. All rights reserved.
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