Functional similarities of human and chicken apolipoprotein A-I: dependence on secondary and tertiary rather than primary structure.
To investigate the sequence requirements for apolipoprotein (apo) AI functions, comparisons of human and chicken apoAI were performed. In lipid binding assays, chicken apoAI was capable of transforming phospholipid vesicles into discoidal bilayer structures, similar in both size and apolipoprotein content to those produced with human apoAI under the same conditions. Human and chicken apoAI were indistinguishable in their relative abilities to prevent phospholipase C-induced aggregation of human low density lipoprotein. This activity, which is dependent upon formation of a stable interaction with the modified lipoprotein, represents a sensitive measure of apolipoprotein association with spherical lipoprotein particles. The ability of chicken versus human apoAI to mobilize the regulatory pool of cholesterol available for esterification by acyl-CoA:cholesterol acyltransferase by human fibroblasts was also assessed. Lipid-free chicken and human apoAI were equivalent in their ability to deplete cholesterol from this pool, as were intact chicken high density lipoprotein (HDL) and human HDL(3). Based on the overall sequence identity of chicken and human apoAI (48%), and comparison of regions thought to be responsible for key apoAI functions, these data indicate that amphipathic alpha-helical structure, rather than specific amino acid sequence, is the major determinant of apoAI lipid binding and ability to mobilize the regulatory pool of cellular cholesterol.
Kiss
R S
RS
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Ryan
R O
RO
Francis
G A
GA
D000595:Amino Acid Sequence; D000818:Animals; D016632:Apolipoprotein A-I / Q000737:chemistry / Q000494:pharmacology*; D002478:Cells, Cultured; D002645:Chickens; D002784:Cholesterol / Q000032:analysis / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry; D008075:Lipoproteins, HDL / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000378:metabolism; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000737:chemistry; D011487:Protein Conformation; D016415:Sequence Alignment; D010738:Type C Phospholipases
D016632:Apolipoprotein A-I; D008051:Lipid Bilayers; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D010743:Phospholipids; D002784:Cholesterol; D010738:Type C Phospholipases