title: Measurement of glyoxalase gene expression.
abstract: The glyoxalase system is an important component of the enzymatic defence against glycation, preventing particularly quantitatively and functionally important glycation of protein and DNA by methylglyoxal. Expression of genes encoding Glo1 (glyoxalase I) and Glo2 (glyoxalase II) may be induced or suppressed, and rates of proteolysis of Glo1 and Glo2 proteins may change in health and disease. Quantitative assessment of glyoxalase gene expression at the mRNA and protein levels has become a key part of glyoxalase system characterization. For mRNA, there is the common technique of real-time RT (reverse transcription)-PCR and direct quantification of mRNA copy number by the Nanostring™ method. For glyoxalase protein quantification, there is the commonly used Western blotting, and also immunoassay and, in proteome-wide studies, quantitative proteomics and proteome dynamics. We provide protocols for the common methods below and briefly review their application.
authors.affiliation: *Clinical Sciences Research Laboratories, Warwick Medical School, University of Warwick, University Hospital, Coventry CV2 2DX, U.K.
authors.forename: Mingzhan
authors.initials: M
authors.lastname: Xue
authors.affiliation: *Clinical Sciences Research Laboratories, Warwick Medical School, University of Warwick, University Hospital, Coventry CV2 2DX, U.K.
authors.forename: Naila
authors.initials: N
authors.lastname: Rabbani
authors.affiliation: *Clinical Sciences Research Laboratories, Warwick Medical School, University of Warwick, University Hospital, Coventry CV2 2DX, U.K.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Thornalley
mesh_terms: D000818:Animals; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007791:Lactoylglutathione Lyase / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000235:genetics / Q000378:metabolism; D040901:Proteomics; D011765:Pyruvaldehyde / Q000378:metabolism
chemical_list: D020543:Proteome; D011765:Pyruvaldehyde; D007791:Lactoylglutathione Lyase