Proteomic analysis of human low-density lipoprotein reveals the presence of prenylcysteine lyase, a hydrogen peroxide-generating enzyme.
The molecular mechanisms underlying the relationship between low-density lipoprotein (LDL) and the risk of atherosclerosis are not clear. Therefore, detailed information on the protein composition of LDL may help to reveal its role in atherogenesis. Liquid-phase IEF has been used to resolve LDL proteins into well-defined fractions on the basis of pI, which improves the subsequent detection and resolution of low abundance proteins. Besides known LDL-associated proteins, this approach revealed the presence of proteins not previously described to reside in LDL, including prenylcysteine lyase (PCL1), orosomucoid, retinol-binding protein, and paraoxonase-1. PCL1, an enzyme crucial for the degradation of prenylated proteins, generates free cysteine, isoprenoid aldehyde and hydrogen peroxide. Addition of the substrate farnesylcysteine to lipoprotein resulted in a time-dependent generation of H(2)O(2) which was stronger in very low density lipoprotein (VLDL) than in LDL or HDL, reflecting the greater protein content of PCL1 in VLDL. Farnesol, a dead end inhibitor of the PCL1 reaction, reduced H(2)O(2) generation by VLDL. PCL1 is generated along with nascent lipoprotein, as shown by its presence in the lipoprotein secreted by HepG2 cells. The finding that an enzyme associated with atherogenic lipoproteins can itself generate an oxidant suggests that PCL1 may play a significant role in atherogenesis.
Banfi
Cristina
C
Monzino Cardiologic Center IRCCS, Milan, Italy. cristina.banfi@unimi.it
Brioschi
Maura
M
Barcella
Simona
S
Wait
Robin
R
Begum
Shajna
S
Galli
Sabrina
S
Rizzi
Andrea
A
Tremoli
Elena
E
D000328:Adult; D050197:Atherosclerosis / Q000209:etiology; D013437:Carbon-Sulfur Lyases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005591:Chemical Fractionation / Q000379:methods; D005204:Farnesol / Q000378:metabolism; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008074:Lipoproteins / Q000032:analysis* / Q000378:metabolism; D008075:Lipoproteins, HDL / Q000032:analysis / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000032:analysis / Q000378:metabolism; D008079:Lipoproteins, VLDL / Q000032:analysis / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry
D008074:Lipoproteins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008079:Lipoproteins, VLDL; D005204:Farnesol; D006861:Hydrogen Peroxide; C108083:prenylcysteine lyase; D013437:Carbon-Sulfur Lyases