Apolipoprotein B-100 of plasma low density lipoproteins undergoes proteolysis by contact activation factors when plasma is treated with dextran sulfate-500-MgCl2.
Human plasma low density lipoproteins (LDL) isolated by ultracentrifugation showed a single band corresponding to apolipoprotein B-100 (apoB-100) by SDS-gradient gel electrophoresis (GGE). In turn, apoB-100 of LDL precipitated from plasma by dextran sulfate-500 (DS)-MgCl2 exhibited several bands indicative of a degradative process. The degradation was more extensive at 0 degrees C than at either 23 degrees C or 37 degrees C, and appeared to be related to a protease activity that cleaved both the synthetic peptide, Z-Phe-Arg-7-amido-4-methylcoumarin (Z-Phe-Arg-AMC) and apoB-100. Proteolysis was proportional to the DS added to the plasma, was prevented by the kallikrein inhibitor, D-Phe-L-Phe-L-Arg-CHCl2, and was significantly decreased in plasma specimens of patients with either factor XII or prekalikrein deficiency. LDL pre-purified by ultracentrifugation and then precipitated by DS in the absence of plasma exhibited no proteolysis. However, proteolysis was observed when LDL interacted with kallikrein. The two main apolipoproteins of HDL3, apoA-I and apoA-II, were not affected by this proteolytic process. We interpret the results to indicate that the negatively charged surface provided by DS accelerates in plasma the autoactivation of factor XII and the activation of prekallikrein, resulting in an increase of the effective concentration of kallikrein and possibly other proteases and proteolysis of LDL-apoB-100. The higher degree of the DS-induced proteolysis of apoB-100 at 0 degrees C than at 23 degrees C is likely the consequence of enhanced autoactivation of factor XII and a decreased efficiency of plasma inhibitors, such as C1-inhibitor. We speculate that the proteolysis of apoB-100 induced by DS is not limited to this polyanion, but may also be the property of other negatively charged agents, particularly at cold temperatures.
Byrne
R E
RE
Department of Medicine, University of Chicago, Pritzker School of Medicine, IL 60637.
Scanu
A M
AM
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000097:blood*; D016264:Dextran Sulfate; D003911:Dextrans; D005174:Factor XII / Q000032:analysis; D006801:Humans; D008077:Lipoproteins, LDL / Q000097:blood*; D008274:Magnesium; D015636:Magnesium Chloride; D011288:Prekallikrein / Q000032:analysis; D012697:Serine Endopeptidases / Q000378:metabolism; D013696:Temperature; D014461:Ultracentrifugation
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D003911:Dextrans; D008077:Lipoproteins, LDL; D015636:Magnesium Chloride; D005174:Factor XII; D016264:Dextran Sulfate; D011288:Prekallikrein; D012697:Serine Endopeptidases; D008274:Magnesium