title: Effect of phospholipase A2 digestion on the conformation and lysine/fibrinogen binding properties of human lipoprotein[a].
abstract: In vitro hydrolysis of human lipoprotein[a] (Lp[a]) by phospholipase A2 (PLA2) decreased the phosphatidylcholine (PC) content by 85%, but increased nonesterified fatty acids 3.2-fold and lysoPC 12.9-fold. PLA2-treated Lp[a] had a decreased molecular weight, increased density, and greater electronegativity on agarose gels. In solution, PLA2-Lp[a] was a monomer, and when assessed by sedimentation velocity it behaved like untreated Lp[a], in that it remained compact in NaCl solutions but assumed the extended form in the presence of 6-amino hexanoic acid, which was shown previously to have an affinity for the apo[a] lysine binding site II (LBS II) comprising kringles IV5-8. We interpreted our findings to indicate that PLA2 digestion had no effect on the reactivity of this site. This conclusion was supported by the results obtained from lysine Sepharose and fibrinogen binding experiments, in the presence and absence of Tween 20, showing that phospholipolysis had no effect on the reactivity of the LBS-II domain. A comparable binding behavior was also exhibited by the free apo[a] derived from each of the two forms of Lp[a]. We did observe a small increase in affinity of PLA2-Lp[a] to lysine Sepharose and attributed it to changes in reactivity of the LBS I domain (kringle IV10) induced by phospholipolysis. In conclusion, the extensive modification of Lp[a] caused by PLA2 digestion had no significant influence on the reactivity of LBS II, which is the domain involved in the binding of apo[a] to fibrinogen and apoB-100. These results also suggest that phospholipids do not play an important role in these interactions.
authors: Department of Medicine, University of Chicago, 5841 South Maryland Avenue, Chicago, IL 60637, USA.
authors: G M
authors: GM
authors: Fless
authors: E W
authors: EW
authors: Kirk
authors: O
authors: O
authors: Klezovitch
authors: J Y
authors: JY
authors: Santiago
authors: C
authors: C
authors: Edelstein
authors: J
authors: J
authors: Hoover-Plow
authors: A M
authors: AM
authors: Scanu
mesh_terms: D015119:Aminocaproic Acid / Q000494:pharmacology; D002499:Centrifugation, Density Gradient; D005230:Fatty Acids, Nonesterified / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D017270:Lipoprotein(a) / Q000737:chemistry* / Q000378:metabolism*; D008970:Molecular Weight; D010713:Phosphatidylcholines / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011487:Protein Conformation; D012685:Sepharose / Q000031:analogs & derivatives* / Q000378:metabolism; D012965:Sodium Chloride; D012996:Solutions
chemical_list: D005230:Fatty Acids, Nonesterified; D017270:Lipoprotein(a); D010713:Phosphatidylcholines; D012996:Solutions; C030675:lysine-sepharose; D012965:Sodium Chloride; D005340:Fibrinogen; D012685:Sepharose; D010741:Phospholipases A; D054467:Phospholipases A2; D015119:Aminocaproic Acid