Oxidative events cause degradation of apoB-100 but not of apo[a] and facilitate enzymatic cleavage of both proteins

J Lipid Res. 2001 Oct;42(10):1664-70.

Abstract

Lipoprotein [a] (Lp[a]) contains equimolar amounts of apoB-100 and apolipoprotein [a] (apo[a]). Both proteins are amenable to degradation in vivo by mechanisms yet to be clearly defined. In this study, we examined the in vitro susceptibility of LDL and Lp[a], obtained from the same donor, to oxidation by either Cu(2)+ or the combined Crotalus adamanteus phospholipase A2 and soybean lipoxygenase system, monitoring the course of the reaction by the generation of conjugated dienes and fatty acids. In some experiments, treatment with leukocyte elastase (LE) or matrix metalloproteinase 12 (MMP-12) was administered before and after the oxidative step. In the case of Lp[a] we found that with both oxidizing systems, conditions that caused the breakdown of apoB-100 did not degrade apo[a] although oxidation-mediated changes were detected in the latter by intrinsic tryptophan fluorescence spectroscopy. Similar results were obtained with a reassembled Lp[a] obtained by incubating free apo[a] with LDL. Both apo[a] and apoB-100 were cleaved by LE and MMP-12 but the enzymatic cleavage was more marked when the preoxidized proteins were used as a substrate. Taken together, our in vitro studies indicate that apo[a] but not apoB-100 resists oxidative fragmentation, whereas both proteins are cleaved by enzymes of the serine and metalloproteinase families. We speculate that the fragments of apo[a] observed in vivo may be preferentially generated by proteolytic rather than oxidative events, whereas apoB-100 can be degraded by both mechanisms.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Apolipoprotein B-100
  • Apolipoproteins A / metabolism*
  • Apolipoproteins B / metabolism*
  • Blotting, Western
  • Copper Sulfate / metabolism
  • Humans
  • Lipoprotein(a) / metabolism
  • Lipoproteins, LDL / metabolism
  • Lipoxygenase / metabolism
  • Matrix Metalloproteinase 12
  • Metalloendopeptidases / metabolism
  • Oxidants / metabolism*
  • Oxidation-Reduction
  • Pancreatic Elastase / metabolism
  • Peptide Hydrolases / metabolism*
  • Phospholipases A / metabolism
  • Phospholipases A2
  • Spectrometry, Fluorescence
  • Time Factors

Substances

  • Apolipoprotein B-100
  • Apolipoproteins A
  • Apolipoproteins B
  • Lipoprotein(a)
  • Lipoproteins, LDL
  • Oxidants
  • Lipoxygenase
  • Phospholipases A
  • Phospholipases A2
  • Peptide Hydrolases
  • Pancreatic Elastase
  • Metalloendopeptidases
  • MMP12 protein, human
  • Matrix Metalloproteinase 12
  • Copper Sulfate