Primary structure comparison of the proposed low density lipoprotein (LDL) receptor binding domain of human and pig apolipoprotein B: implications for LDL-receptor interactions.
Apolipoprotein B (apoB) is the predominant protein in low density lipoprotein (LDL) and is responsible for LDL binding to the LDL receptor. Although the primary amino acid sequence of human apoB has been determined, little is known about the structural domains involved in mediating apoB binding to the LDL receptor. Amino acid sequence comparisons across species lines provide a means of defining structures that are essential for function. We have sequenced a l.l kb fragment of pig apoB genomic DNA, corresponding to a 363 amino acid segment proposed to mediate human apoB binding to the LDL receptor. In human apoB this domain contains two regions enriched in positively charged amino acids flanking two disulfide-linked cysteine residues. The pig amino acid sequence shared 72% identity with the human sequence. However, there were differences that have significant structural and functional implications. Human apoB arginine-3,359 corresponds to a critical arginine (position 142) residue in the apoE LDL receptor binding domain. In the pig, this arginine residue was not conserved. Also, the two disulfide-linked cysteine residues found near the proposed apoB binding domain were not conserved in the pig. Despite these differences, pig LDL had a higher affinity than human LDL for both the pig and human LDL receptor. Thus, these features are not required for high affinity binding of pig LDL to the LDL receptor, and may not be necessary for the binding of human LDL to the LDL receptor.
Ebert
D L
DL
Department of Biochemistry, University of Wisconsin-Madison 53706.
Maeda
N
N
Lowe
S W
SW
Hasler-Rapacz
J
J
Rapacz
J
J
Attie
A D
AD
D000595:Amino Acid Sequence; D000818:Animals; D001055:Apolipoproteins B / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D001057:Apolipoproteins E / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D008969:Molecular Sequence Data; D010641:Phenotype; D011973:Receptors, LDL / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity; D013329:Structure-Activity Relationship; D013552:Swine
D001055:Apolipoproteins B; D001057:Apolipoproteins E; D008077:Lipoproteins, LDL; D011973:Receptors, LDL