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Enzymatic assessment of paraoxonase 1 activity on HDL subclasses: a practical zymogram method to assess HDL function.
- Source :
-
Clinica chimica acta; international journal of clinical chemistry [Clin Chim Acta] 2013 Jan 16; Vol. 415, pp. 162-8. Date of Electronic Publication: 2012 Oct 30. - Publication Year :
- 2013
-
Abstract
- Background: We developed a practical method for analysis of PON-1 enzymatic activity in HDL subclasses.<br />Methods: The assay uses 4-12% polyacrylamide gradient gels, phenylacetate as substrate coupled with densitometric phenol detection using 4-aminoantipyrine. The measurement PON-1 activity in situ across the HDL subclasses has a strong correlation with the kinetic microplate assay for total PON-1 activity, r=0.91, p<0.001.<br />Results: The same HDL-C level, healthy subjects (n=33) display a large difference in the ratio of PON-1 activity in small vs. large HDL. Since PON-1 activity is larger in HDL(3) we propose that this difference has a potent predictive value for clinical risk assessment and therapeutic choice. Our method also offers the advantage of assessing the distribution of up to six different HDL apolipoproteins in the same gel after transfer.<br />Conclusions: We seek to further dissect the cause of a different distribution of PON-1 activity in HDL subclasses by employing this method that permits practical, inexpensive analysis of antioxidant function of HDL subclasses and has the potential for application in clinical chemistry and to shed some light on the importance of PON-1 distribution.<br /> (Copyright © 2012 Elsevier B.V. All rights reserved.)
- Subjects :
- Adult
Ampyrone chemistry
Aryldialkylphosphatase metabolism
Electrophoresis, Polyacrylamide Gel
Female
Humans
Lipoproteins, HDL chemistry
Lipoproteins, HDL classification
Lipoproteins, HDL isolation & purification
Male
Middle Aged
Phenol analysis
Phenylacetates chemistry
Aryldialkylphosphatase analysis
Enzyme Assays
Lipoproteins, HDL metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1873-3492
- Volume :
- 415
- Database :
- MEDLINE
- Journal :
- Clinica chimica acta; international journal of clinical chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 23122967
- Full Text :
- https://doi.org/10.1016/j.cca.2012.10.044