부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ab.2008.09.039 Byline: G. Hoffner (a), G. van der Rest (b), P.M. Dansette (c), P. Djian (a) Keywords: [N.sup.I[micro] -([gamma]-Glutamyl)] lysine isodipeptide; Transglutaminase; Neurodegenerative diseases; Huntington disease; Quantitation method; Liquid chromatography; Mass spectrometry Abstract: Transglutaminases catalyze the formation of N.sup.I[micro] -([gamma]-glutamyl) isodipeptide crosslinks between proteins. These enzymes are thought to participate in a number of diseases, including neurological disease and cancer. A method associating liquid chromatography and multiple stage mass spectrometry has been developed for the simultaneous quantitation of [N.sup.I[micro] -([gamma]-glutamyl) lysine] isodipeptide and lysine on an ion trap mass spectrometer. Highly specific detection has been achieved in MS.sup.3 mode. The method includes a derivatization step consisting of butylation of carboxylic groups and acetylation of amide groups, a liquid-liquid extraction, and a 19-min separation on a 100x2.1-mm Beta-basic C.sub.18 column with an acetonitrile gradient elution..sup.13C.sub.6-.sup.15N.sub.2 isotopes of the isodipeptide and the lysine serve as internal standards. The assay was linear in the range of 50pmol/ml to 75nmol/ml for the isodipeptide and the range of 10nmol/ml to 3.5[mu]mol/ml for the lysine, with correlation coefficients greater than 0.99 for both ions. Intra- and inter-day coefficients of variation ranged from 3.5 to 15.9%. The method was successfully applied to human biological samples known to be crosslinked by transglutaminase such as cornified envelopes of epidermis, fibrin, and normal and Huntington disease brain. Author Affiliation: (a) Laboratoire Regulation de la Transcription et Maladies Genetiques, UPR 2228 CNRS, Institut Interdisciplinaire des Sciences du Vivant des Saints-Peres, Universite Paris Descartes, 75270 Paris Cedex 06, France (b) Laboratoire des Mecanismes Reactionnels, UMR 7651 CNRS, Departement de Chimie, Ecole Polytechnique, 91128 Palaiseau Cedex, France (c) Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, UMR 8601 CNRS, Institut Interdisciplinaire des Sciences du Vivant des Saints-Peres, Universite Paris Descartes, 75270 Paris Cedex 06, France Article History: Received 4 July 2008