Mutation of lysine residues in apolipoprotein B-100 causes defective lipoprotein[a] formation

• Main Authors: Catherine Y.Y. Liu, Ric Broadhurst, Santica M. Marcovina, Sally P.A. McCormick
• Format: Article
• Language: English
• Published: Elsevier 2004-01-01
• Series: Journal of Lipid Research
• Subjects: apolipoprotein[a]; assembly; coronary heart disease; lysine; site-directed mutagenesis; transgenic mice
• Online Access: http://www.sciencedirect.com/science/article/pii/S0022227520319210

Lipoprotein[a] (Lp[a]) is assembled by a two-step process involving an initial lysine-dependent binding between apolipoprotein B-100 (apoB-100) and apolipoprotein[a] (apo[a]) that facilitates the formation of a disulphide bond between apoB-100Cys4,326 and apo[a]Cys4,057. Previous studies of transgenic mice expressing apoB-95 (4,330 amino acids) and apoB-97 (4,397 amino acids) have shown that apoB-100 amino acids 4,330–4,397 are important for the initial binding to apo[a]. Furthermore, a lysine-rich peptide spanning apoB-100 amino acids 4,372–4,392 has recently been shown to bind apo[a] and inhibit Lp[a] assembly in vitro. This suggests that a putative apo[a] binding site exists in the apoB-4,372–4,392 region. The aim of our study was to establish whether the apoB-4,372–4,392 sequence was important for Lp[a] assembly in the context of the full-length apoB-100. Transgenic mice were created that expressed a mutant human apoB-100, apoB-100K4→S4, in which all four lysine residues in the 4,372–4,392 sequence were mutated to serines. The apoB-100K4→S4 mutant showed a reduced capacity to form Lp[a] in vitro compared with wild-type human apoB-100. Double transgenic mice expressing both apoB-100K4→S4 and apo[a] contained significant amounts of free apo[a] in the plasma, indicating a less-efficient assembly of Lp[a] in vivo.Taken together, these results clearly show that the apoB-4,372–4,392 sequence plays a role in Lp[a] assembly.