title: Enzymatic assessment of paraoxonase 1 activity on HDL subclasses: a practical zymogram method to assess HDL function.
abstract: BACKGROUND
  We developed a practical method for analysis of PON-1 enzymatic activity in HDL subclasses.
  
  
  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.
  
  
  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.
  
  
  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.
authors.affiliation: Glycation, Oxidation and Disease Laboratory, Touro University-California, Vallejo, CA 94592, USA. alejandro.gugliucci@tu.edu
authors.forename: Alejandro
authors.initials: A
authors.lastname: Gugliucci
authors.forename: Russell
authors.initials: R
authors.lastname: Caccavello
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Kotani
authors.forename: Naoki
authors.initials: N
authors.lastname: Sakane
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kimura
mesh_terms: D000328:Adult; D000675:Ampyrone / Q000737:chemistry; D043303:Aryldialkylphosphatase / Q000032:analysis* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays; D005260:Female; D006801:Humans; D008075:Lipoproteins, HDL / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D019800:Phenol / Q000032:analysis; D010648:Phenylacetates / Q000737:chemistry
chemical_list: D008075:Lipoproteins, HDL; D010648:Phenylacetates; D000675:Ampyrone; D019800:Phenol; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human; C025136:phenylacetic acid