| Abstract: | Lipoproteina] (Lpa]) is assembled by a two-step process involving an initial lysine-dependent binding between apolipoprotein B-100 (apoB-100) and apolipoproteina] (apoa]) that facilitates the formation of a disulphide bond between apoB-100Cys4,326 and apoa]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 apoa]. Furthermore, a lysine-rich peptide spanning apoB-100 amino acids 4,372-4,392 has recently been shown to bind apoa] and inhibit Lpa] assembly in vitro. This suggests that a putative apoa] 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 Lpa] 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 Lpa] in vitro compared with wild-type human apoB-100. Double transgenic mice expressing both apoB-100K4-->S4 and apoa] contained significant amounts of free apoa] in the plasma, indicating a less-efficient assembly of Lpa] in vivo. Taken together, these results clearly show that the apoB-4,372-4,392 sequence plays a role in Lpa] assembly. |
