HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Papers In Press, published online ahead of print July 1, 2007
J. Lipid Res., doi:10.1194/jlr.D600045-JLR200

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: D600045-JLR200v1 48/7/1637    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gaidukov, L. Articles by Tawfik, D. S. Search for Related Content PubMed PubMed Citation Articles by Gaidukov, L. Articles by Tawfik, D. S. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol. 48, 1637-1646, July 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Methods

The development of human sera tests for HDL-bound serum PON1 and its lipolactonase activity

Leonid Gaidukov and Dan S. Tawfik¹

Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of one table and five figures.

Published, JLR Papers in Press, April 13, 2007.

¹ To whom correspondence should be addressed. e-mail: tawfik{at}weizmann.ac.il

Serum paraoxonase (PON1) is a lipolactonase that associates with HDL-apolipoprotein A-I (HDL-apoA-I) and thereby plays a role in the prevention of atherosclerosis. Current sera tests make use of promiscuous substrates and provide no indications regarding HDL-PON1 complex formation. We developed new enzymatic tests that detect total PON1 levels, irrespective of HDL status and R/Q polymorphism, as well as the degree of catalytic stimulation and increased stability that follow PON1's tight binding to HDL-apoA-I. The tests are based on measuring total PON1 levels with a fluorogenic phosphotriester, measuring the lipolactonase activity with a chromogenic lactone, and assaying the enzyme's chelator-mediated inactivation rate. The latter two are affected by tight HDL binding and thereby derive the levels of the serum PON1-HDL complex. We demonstrate these new tests with a group of healthy individuals (n = 54) and show that the levels of PON1-HDL vary by a factor of 12. Whereas the traditionally applied paraoxonase and arylesterase tests weakly reflect PON1-HDL levels (R = 0.64), the lipolactonase test provides better correlation (R = 0.80). These new tests indicate the levels and activity of PON1 in a physiologically relevant context as well as the levels and quality of the HDL particles with which the enzyme is associated.

Supplementary key words blood tests • lactones • lipoproteins • high density lipoprotein-bound enzymes • enzyme stability • atherosclerosis • paraoxonase

Abbreviations: apoA-I, apolipoprotein A-I; DEPCyMC, 7-O-diethyl phosphoryl 3-cyano 4-methyl 7-hydroxycoumarin; DTNB, 5,5'-dithio-bis-2-nitrobenzoic acid; PON1, serum paraoxonase; TBBL, 5-thiobutyl butyrolactone

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

This article has been cited by other articles:

				P. W. Connelly, B. Zinman, G. F. Maguire, M. Mamakeesick, S. B. Harris, R. A. Hegele, R. Retnakaran, and A. J. G. Hanley Association of the novel cardiovascular risk factors paraoxonase 1 and cystatin C in type 2 diabetes J. Lipid Res., June 1, 2009; 50(6): 1216 - 1222. [Abstract] [Full Text] [PDF]

				P. W. Connelly Increased serum advanced glycation end products are associated with impairment in HDL antioxidative capacity in diabetic nephropathy Nephrol. Dial. Transplant., August 1, 2008; 23(8): 2699 - 2700. [Full Text] [PDF]

				B. Thyagarajan, D. R. Jacobs Jr., J. J. Carr, O. Alozie, M. W. Steffes, P. Kailash, J. H. Hayes, and M. D. Gross Factors Associated with Paraoxonase Genotypes and Activity in a Diverse, Young, Healthy Population: The Coronary Artery Risk Development in Young Adults (CARDIA) Study Clin. Chem., April 1, 2008; 54(4): 738 - 746. [Abstract] [Full Text] [PDF]