Measurement of glyoxalase activities

• DOI: 10.1042/bst20140010
• Publication Date: 2014-03-20T12:39:45Z
• Authors: Makoto Arai; Naomi Nihonmatsu-...; Masanari Itokawa; Naila Rabbani; Paul J. Thornalley
• Keywords: Lactoylglutathione lyase

Glyoxalase I catalyses the isomerization of hemithioacetal formed non-enzymatically from methylglyoxal and glutathione to S-D-lactoylglutathione. The activity glyoxalase is conventionally measured spectrophotometrically by following increase in A240 for which change molar absorption coefficient Δε240=2.86 mM⁻¹·cm⁻¹. pre-formed situ incubation 50 mM sodium phosphate buffer (pH 6.6) at 37°C 10 min. cell extract then added, monitored over 5 min, initial rate hence deduced with correction blank. given units per mg protein or number where one unit amount enzyme that formation 1 μmol S-D-lactoylglutathione min under assay conditions. II hydrolysis D-lactate glutathione. also decrease Δε240=-3.10 It measurements have been modified use a UV-transparent microplate higher sample throughput.