Title of article :
Immunohistochemical detection of N-homocysteinylated proteins in humans and mice
Author/Authors :
Joanna Per?a-Kajan، نويسنده , , Olaf Stanger، نويسنده , , Micha? ?uczak، نويسنده , , Agnieszka Zio?kowska، نويسنده , , Ludwik K. Malendowicz، نويسنده , , Tomasz Twardowski، نويسنده , , Sarka Lhotak، نويسنده , , Richard C. Austin، نويسنده , , Hieronim Jakubowski، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2008
Pages :
7
From page :
473
To page :
479
Abstract :
N-homocysteinylation of -amino group of protein lysine residues by homocysteine (Hcy) thiolactone has been implicated in vascular disease in humans. We have previously generated polyclonal rabbit anti-N-Hcy-protein IgG antibodies that specifically recognize the N -Hcy-Lys epitope on N-homocysteinylated proteins. The present work was undertaken to examine the utility of these antibodies for the immunohistochemical detection of N-homocysteinylated proteins in biological samples. We found that the rabbit antibody specifically detected N-Hcy-protein in a dot-blot assay, that the signal resulting from the reaction of the antibody with N-Hcy-protein depended on the amount of the antigen, and that the sensitivity of the assay was protein-dependent. The rabbit anti-N-Hcy-protein IgG also specifically detected N -Hcy-Lys epitopes in human tissues, as shown by positive immunohistochemical staining of myocardium and aorta samples from cardiac surgery patients, and a lack of staining when the antibody was pre-adsorbed with N-Hcy-albumin. We also observed increased immunohistochemical staining for N-Hcy-proteins in aortic lesions from ApoE−/− mice with hyperhomocysteinemia induced by a high methionine diet, relative to ApoE−/− mice fed a control chow diet. In conclusion, polyclonal rabbit anti-N-Hcy-protein antibody can detect and monitor N-homocysteinylated proteins in human and mouse tissues with good sensitivity and specificity.
Keywords :
atherosclerosis , antibodies , Homocysteine thiolactone , N-homocysteinylated protein
Journal title :
Biomedicine and Pharmacotherapy
Serial Year :
2008
Journal title :
Biomedicine and Pharmacotherapy
Record number :
478125
Link To Document :
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