ChE, GST and CAT: evaluation of the efficiency of a combined buffer for protein extraction.
Toxicity and biotransformation of several earthworm contaminants are widely evaluated nowadays using biochemical biomarkers. Many investigations track enzyme activities as biomarkers of neurotoxicity (cholinesterase (ChE)), metabolisation (glutathione-S-transferase (GST)) and oxidative stress (catalase (CAT)). This study proposes an evaluation of the use of a combined buffer, to extract proteins from earthworms and then analyse the 3 biomarkers. The method provides good results and allows protein extraction and quantitative determination of biomarkers with the same efficiency as the enzyme-specific buffers. It decreases preparation time and permits a study of the biomarkers on the same individual with only one homogenisation.
Schreck
E
E
Laboratoire Chimie et Biochimie des Interactions, Université de Toulouse-Centre Universitaire JF Champollion, Albi, France. eva.schreck@univ-jfc.fr
Geret
F
F
Gontier
L
L
Treilhou
M
M
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002021:Buffers; D002374:Catalase / Q000302:isolation & purification*; D002802:Cholinesterases / Q000302:isolation & purification*; D005982:Glutathione Transferase / Q000302:isolation & purification*; D007700:Kinetics; D009835:Oligochaeta / Q000187:drug effects / Q000201:enzymology*; D012989:Soil Pollutants / Q000633:toxicity; D014020:Tissue Extracts / Q000737:chemistry
D015415:Biomarkers; D002021:Buffers; D012989:Soil Pollutants; D014020:Tissue Extracts; D002374:Catalase; D005982:Glutathione Transferase; D002802:Cholinesterases