Skip Navigation


Journal of Biochemistry Advance Access originally published online on January 29, 2007
Journal of Biochemistry 2007 141(4):495-502; doi:10.1093/jb/mvm050
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
141/4/495    most recent
mvm050v2
mvm050v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Hitomi, Y. H.
Right arrow Articles by Ogino, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hitomi, Y. H.
Right arrow Articles by Ogino, K.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2007 The Japanese Biochemical Society.

Disposition of Protein-bound 3-nitrotyrosine in Rat Plasma Analysed by a Novel Protocol for HPLC–ECD

Yoshiaki H. Hitomi1, Junna Okuda1, Hirohito Nishino2, Yasuhiro Kambayashi1, Yuri Hibino1, Kei Takemoto3, Tomoko Takigawa3, Hideki Ohno4, Naoyuki Taniguchi5 and Keiki Ogino3,*

1Department of Environmental and Preventive Medicine, Graduate School of Medical Science, Kanazawa University, 13-1, Takaramachi, Kanazawa, Ishikawa 920-8640; 2Eicom Corporation, 24-2, Enmenda, Shimotoba, Fushimi-ku, Kyoto 612-8474; 3Department of Public Health, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1, Shikada, Okayama 700-8558; 4Department of Molecular Predictive Medicine and Sport Science, Kyorin University, School of Medicine, 6-20-2, Shinkawa, Mitaka, Tokyo 181-8611; and 5Department of Disease Glycomics, Research Institute for Microbial Diseases, Osaka University, 3-1, Yamadaoka, Suita, Osaka 565-0871, Japan

*To whom correspondence should be addressed. Tel: +81-86-235-7184, Fax: +81-86-226-0715, E-mail: kogino{at}md.okayama-u.ac.jp

Received November 30, 2006; Accepted January 24, 2007


  Abstract

3-Nitrotyrosine (NTyr) is considered as a biomarker of the generation of reactive nitrogen species (RNS). However, it is still difficult to determine its concentration in biological samples. To develop a reliable and high-throughput method, we optimized the conditions for high performance liquid chromatography and electrochemical detection (HPLC–ECD). The best separation of NTyr was achieved using a highly acidic mobile phase (pH 2.5). The concentration of protein-bound NTyr in plasma protein was 593.6 ± 53.8 fmol/mg in rats treated with lipopolysaccharide (LPS) and 114.4 ± 27.6 fmol/mg in control. After intravenous administration of in vitro-nitrated plasma protein, NTyr concentration decreased; the half-life was 63.4 ± 16.8 h. Consistently, protein-bound NTyr concentration in plasma after LPS treatment declined gradually, but was detectable for 1 week. Our protocol is reproducible and suitable for analysing multiple clinical samples to study RNS production in vivo.

Key Words: electrochemical detection, HPLC, lipopolysaccharide, nitric oxide, 3-nitrotyrosine

Abbreviations: ECD, electrochemical detection; GS/MS, gas chromatography/mass spectrometry; HPLC, high performance liquid chromatography; LC/MS, liquid chromatography/mass spectrometry; LPS, lipopolysaccharide; NO, nitric oxide; NTyr, 3-nitrotyrosine; PBS, phosphate buffered saline; PVDF, polyvinylidene difluoride; RNS, reactive nitrogen species; TBST, Tris-buffered saline with Tween 20


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.