Fluorogenic tagging of protein 3-nitrotyrosine with 4-(aminomethyl)benzenesulfonate (ABS) in tissues: a useful alternative to immunohistochemistry for fluorescence microscopy imaging of protein nitration
dc.contributor.author | Sharov, Victor S. | |
dc.contributor.author | Pal, Ranu | |
dc.contributor.author | Dremina, Elena S. | |
dc.contributor.author | Michaelis, Elias K. | |
dc.contributor.author | Schoneich, Christian | |
dc.date.accessioned | 2017-03-09T21:30:12Z | |
dc.date.available | 2017-03-09T21:30:12Z | |
dc.date.issued | 2013-11-15 | |
dc.identifier.citation | Sharov, V.s., R. Pal, E.s. Dremina, E.k. Michaelis, and C. Schoneich. "Fluorogenic Tagging of Protein 3-nitrotyrosine with 4-(aminomethyl) benzene Sulfonate in Tissues: A Useful Alternative to Immunohistochemistry for Fluorescence Microscopy Imaging of Protein Nitration." Free Radical Biology and Medicine 53.10 (2012): 1877-885. | en_US |
dc.identifier.uri | http://hdl.handle.net/1808/23389 | |
dc.description.abstract | Protein tyrosine nitration is a common biomarker of biological aging and diverse pathologies associated with the excessive formation of reactive oxygen and nitrogen species. Recently, we suggested a novel fluorogenic derivatization procedure for the detection of 3-nitrotyrosine (3-NT) using benzylamine derivatives to convert specifically protein or peptide bound 3-NT to a highly fluorescent benzoxazole product. In the current study, we applied this procedure to fluorogenic derivatization of protein 3-NT in sections from adult rat cerebellum in order to: (i) test this method in imaging nitrated proteins in fixed brain tissue sections, and (ii) compare the chemical approach to immunohistochemical labeling with anti-3-NT antibodies. Immunofluorescence analysis of cerebellar sections using anti-3-NT antibodies showed differential levels of immunostaining in the molecular, Purkinje, and granule cell layers of the cerebellar cortex; in agreement with previous reports, the Purkinje cells were most highly labeled. Importantly, fluorogenic derivatization reactions of cerebellar proteins with 4-(aminomethyl)benzenesulfonic acid (ABS) and K3Fe(CN)6 at pH 9, following sodium dithionite (SDT) reduction of 3-NT to 3-aminotyrosine (3-AT), showed a very similar pattern of relative intensity of cell labeling and improved resolution when compared with antibody labeling. Our data demonstrate that ABS-derivatization may be either a useful alternative or a complimentary approach to immunolabeling in imaging protein nitration in cells and tissues, including under conditions of dual labeling with antibodies to cell proteins, thus allowing for cellular co-localization of nitrated proteins and any protein of interest. | en_US |
dc.publisher | Elsevier | en_US |
dc.rights | This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 3.0 (CC BY-NC-ND 3.0 US), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. | en_US |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/3.0/ | |
dc.subject | Protein nitration | en_US |
dc.subject | 3-nitrotyrosine | en_US |
dc.subject | Cerebellum | en_US |
dc.subject | Purkinje cells | en_US |
dc.subject | 4-(aminomethyl)benzenesulfonate (ABS) | en_US |
dc.subject | Fluorogenic derivatization | en_US |
dc.subject | Fluorescence microscopy | en_US |
dc.subject | Immunohistochemistry | en_US |
dc.subject | Synaptophysin | en_US |
dc.subject | SIN-1 | en_US |
dc.title | Fluorogenic tagging of protein 3-nitrotyrosine with 4-(aminomethyl)benzenesulfonate (ABS) in tissues: a useful alternative to immunohistochemistry for fluorescence microscopy imaging of protein nitration | en_US |
dc.type | Article | en_US |
kusw.kuauthor | Michaelis, Eli K. | |
kusw.kudepartment | Higuchi Biosciences Center | en_US |
dc.identifier.doi | 10.1016/j.freeradbiomed.2012.08.582 | en_US |
kusw.oaversion | Scholarly/refereed, author accepted manuscript | en_US |
kusw.oapolicy | This item meets KU Open Access policy criteria. | en_US |
dc.rights.accessrights | openAccess |
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Except where otherwise noted, this item's license is described as: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 3.0 (CC BY-NC-ND 3.0 US), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.