Is nitrotyrosine generated in human erythrocytes in circulation?

Citation
K. Kikugawa et al., Is nitrotyrosine generated in human erythrocytes in circulation?, BIOL PHAR B, 23(4), 2000, pp. 379-385
Citations number
60
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Pharmacology & Toxicology
Journal title
BIOLOGICAL & PHARMACEUTICAL BULLETIN
ISSN journal
0918-6158 → ACNP
Volume
23
Issue
4
Year of publication
2000
Pages
379 - 385
Database
ISI
SICI code
0918-6158(200004)23:4<379:INGIHE>2.0.ZU;2-S
Abstract
Nitrotyrosine is considered a stable biomarker of reactive nitrogen species , including nitrogen dioxide (NO2) and peroxynitrous acid (ONOOH) in biomat erials. There are inconsistent observations on the detection of free and pr otein-associated nitrotyrosine in normal human plasma. Human erythrocytes, differentiated from erythrocyte precursor cells in the bone marrow, circula ting in the body for an average of 120 d, and finally removed by spleen mac rophages, may be exposed to reactive nitrogen species. In the present study , membrane proteins and hemoglobin from the senescent erythrocyte populatio n were compared with those from young erythrocytes separated from the same individuals in their nitrotyrosine presence using newly prepared rabbit pol yclonal anti-nitrotyrosine-ribonuclease A and anti-nitro(N-butoxycarbonyl)t yrosine-bovine serum albumin antibodies. Sodium dodecyl sulfate-polyacrylam ide gel electrophoresis of the membranes and hemoglobin, and subsequent Wes tern blot analysis, showed that these antibodies only slightly bind to the bands of the proteins from both young and senescent erythrocytes, whereas t hese antibodies definitely bind to the protein bands of membranes and hemog lobin nitrated by NO2 or ONOOH in vitro. This result indicates that nitroty rosine is not detected in the membrane proteins and hemoglobin in human nor mal erythrocytes in circulation. However, this does not conclude that eryth rocytes are not exposed to reactive nitrogen species in the circulation.