Comparison of the inhibitory effects of ApoB100 and tissue factor pathway inhibitor on tissue factor and the influence of lipoprotein oxidation.
The procoagulant activity of tissue factor is regulated by circulating inhibitors such as tissue factor pathway inhibitor (TFPI) and LDL. These 2 inhibitors also readily associate making the distinction between their activities difficult. We have examined the relative contributions of intact and C-terminal truncated TFPI and ApoB100. By following the inhibitory potential of the preparations, over a period of 120 minutes, it was demonstrated that TFPI and LDL-resembling particles inhibited tissue factor at different rates. TFPI was found to be a short, fast-acting inhibitor, whereas the action of LDL-resembling particles was more prolonged but slower. The oxidation of LDL has been closely associated with the development of cardiovascular disease, including atherosclerosis and thrombosis. Positively charged amino acids, particularly lysine residues, are prone to alterations via the formation of adducts by lipid peroxidation products. These residues are important in the inhibition of tissue factor activity by ApoB100. They also play an important role in the inhibitory Kunitz domains of TFPI. We have shown that the decline in the ability of LDL to inhibit tissue factor was as a result of modifications in LDL arising from oxidation. By examining the effects of oxidation on full-length and C-terminal truncated TFPI bound to LDL-resembling particles, we found that TFPI is only affected when in close association with ApoB100. C-terminal truncated TFPI was not affected significantly by oxidation. Finally, chemical modification of lysine and arginine residues reduced the overall inhibition of tissue factor by TFPI. We propose that TFPI and LDL act separately to inhibit tissue factor in vivo. However, the oxidation of LDL can alter both the endogenous activity of ApoB100 and reduce that of closely associated TFPI, compromising normal hemostasis.
Department of Biochemistry and Molecular Biology, Royal Free and University College Medical Schools (Royal Free Campus), London, UK. Camille@RFHSM.AC.UK
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C
Ettelaie
B R
BR
Wilbourn
J M
JM
Adam
N J
NJ
James
K R
KR
Bruckdorfer
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000494:pharmacology*; D003300:Copper / Q000494:pharmacology; D006801:Humans; D008074:Lipoproteins / Q000494:pharmacology*; D008077:Lipoproteins, LDL / Q000378:metabolism* / Q000502:physiology; D010084:Oxidation-Reduction; D011973:Receptors, LDL / Q000378:metabolism; D013329:Structure-Activity Relationship; D013925:Thromboplastin / Q000037:antagonists & inhibitors*
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D008074:Lipoproteins; D008077:Lipoproteins, LDL; D011973:Receptors, LDL; C051928:lipoprotein-associated coagulation inhibitor; C099252:oxidized low density lipoprotein; D003300:Copper; D013925:Thromboplastin