PMID: 6143252Jan 1, 1984

Detection of epsilon(gamma-glutamyl) lysine

Molecular and Cellular Biochemistry
M Griffin, J Wilson


Detection of epsilon(gamma-glutamyl) lysine crosslinks is not only necessary for establishing the importance of the dipeptide as a post-translational modification of proteins, but provides information as to the importance of the transglutaminase enzyme in a biological system. The crosslink may be detected using both indirect and direct methodology. Indirect methods for its detection include measurement of 'masked lysines' within a protein, detection of polymer formation by gel-electrophoresis and the inhibition of crosslinking by the incorporation of small molecular weight amines into the substrate protein. Direct methods for the detection of epsilon(gamma-glutamyl) lysine require the actual isolation of the dipeptide following its release from the sample protein by exhaustive proteolytic digestion. Separation of the dipeptide from other components of the digest may be achieved by either ion-exchange chromatography or gel filtration and its qualitative identification achieved by techniques such as paper-electrophoresis or thin layer chromatography. Quantitative estimation of epsilon(gamma-glutamyl) lysine normally involves its further separation by ion-exchange chromatography and its post-column detection following derivatisati...Continue Reading


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