Apolipoprotein J is associated with paraoxonase in human plasma.
Apolipoprotein J (apoJ)-containing high-density lipoproteins (HDL), isolated from human plasma by immunoaffinity chromatography, are associated with apoAI and a protein of approximately 44 kDa. In order to advance our understanding of apoJ's role in the vasculature, a comprehensive investigation was performed to identify and characterize this 44-kDa protein and to study its interaction with apoJ. The 44-kDa protein, a monomeric glycoyslated polypeptide, was identified by N-terminal sequencing as serum paraoxonase. Paraoxonase exists in two oxidation states: one contains all free cysteines while the other has one disulfide bond between Cys42 and Cys284. Northern analysis of eight human tissues shows paraoxonase message present only in the liver. The majority of apoJ/paraoxonase-HDL are 90-140 kDa; however, not all of the plasma paraoxonase is associated with apoJ. The specificity of the apoJ/paraoxonase interaction, inferred by the constant mole ratio of the two proteins in affinity-purified apoJ-HDL, is confirmed in direct binding assays. For purified proteins, there is more than a 5-fold increase in the apparent affinity of apoJ for immobilized paraoxonase as the paraoxonase coating concentration is increased from 0.5 to 2.0 micrograms/mL. Both oxidation states of paraoxonase bind to apoJ with equal affinity. Our data combined with other evidence suggest that the plasma link of apoJ with paraoxonase will be implicated as a predictor of vascular damage.
Kelso
G J
GJ
Department of Pharmacology and Cell Biophysics, College of Medicine, University of Cincinnati, Ohio 45267.
Stuart
W D
WD
Richter
R J
RJ
Furlong
C E
CE
Jordan-Starck
T C
TC
Harmony
J A
JA
D000595:Amino Acid Sequence; D043303:Aryldialkylphosphatase; D051152:Clusterin; D018076:DNA, Complementary / Q000235:genetics; D004220:Disulfides / Q000737:chemistry; D004950:Esterases / Q000097:blood* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000097:blood* / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes; D008099:Liver / Q000201:enzymology; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D012333:RNA, Messenger / Q000032:analysis; D017421:Sequence Analysis
C495242:CLU protein, human; D051152:Clusterin; D018076:DNA, Complementary; D004220:Disulfides; D006023:Glycoproteins; D007527:Isoenzymes; D018832:Molecular Chaperones; D010446:Peptide Fragments; D012333:RNA, Messenger; D004950:Esterases; D043303:Aryldialkylphosphatase