Advertisement
J. Lipid Res.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

A more recent version of this article appeared on June 1, 2005

Papers In Press, published online ahead of print April 1, 2005
J. Lipid Res., doi:10.1194/jlr.D500005-JLR200
This Article
Right arrow Full Text (Accepted Manuscript)
Right arrow All Versions of this Article:
D500005-JLR200v1
46/6/1339    most recent
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 Download to citation manager
Right arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Sabirsh, A.
Right arrow Articles by Owman, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sabirsh, A.
Right arrow Articles by Owman, C.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Submitted on February 15, 2005
Revised on March 17, 2005
Accepted on March 24, 2005

Fluorescent leukotriene B4: potential applications

Alan Sabirsh, Anders Wetterholm, Jesper Bristulf, Hakon Leffler, Jesper Z. Haeggstrom, and Christer Owman

Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm 17177

Corresponding Author: Alan.Sabirsh{at}mbb.ki.se

Leukotriene B{super4} (LTB{super4}) is a potent lipid mediator of inflammation, which acts primarily via a seven-transmembrane spanning, G-protein coupled, receptor denoted BLT{sub1}. Here we describe the synthesis and characterization of fluorescent analogues of LTB{super4} which are easy to produce, inexpensive and without the disadvantages of a radioligand. Fluorescent LTB{super4} is useful for labelling LTB{super4} receptors for which no antibodies are available and, for performing one-step fluorescence polarisation assays conducive to high-throughput screening. We found that orange and green fluorescent LTB{super4} were full agonists that activated the LTB{super4} receptor BLT{sub1} with EC50s of 68 and 40 nM respectively (4.5 nM for unmodified LTB{super4}). Flow cytometric measurements and confocal imaging showed that fluorescent LTB{super4} co-localised with BLT{sub1}. Fluorescence polarisation measurements showed that orange fluorescent LTB{super4} bound to BLT{sub1} with a Kd of 66 nM, and that this binding could be displaced by unlabelled LTB{super4}, and other BLT{sub1} specific ligands. Fluorescent LTB{super4} analogs were also able to displace tritiated LTB{super4}. Orange fluorescent LTB{super4} binding to EGFP-tagged BLT{sub1} could be observed using fluorescence resonance energy transfer (FRET). In addition to being a useful alternative to radiolabelled LTB{super4}, the unique properties of fluorescently labelled LTB{super4} allow a variety of detection technologies to be employed.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
L. Arcemisbehere, T. Sen, L. Boudier, M.-N. Balestre, G. Gaibelet, E. Detouillon, H. Orcel, C. Mendre, R. Rahmeh, S. Granier, et al.
Leukotriene BLT2 Receptor Monomers Activate the Gi2 GTP-binding Protein More Efficiently than Dimers
J. Biol. Chem., February 26, 2010; 285(9): 6337 - 6347.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
 All ASBMB Journals   Journal of Biological Chemistry 
 Molecular and Cellular Proteomics   ASBMB Today 
Copyright © 2005 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement