The central type Y amphipathic alpha-helices of apolipoprotein AI are involved in the mobilization of intracellular cholesterol depots.
We studied the role of a central domain of human apolipoprotein AI (apoAI) in cholesterol mobilization and removal from cells. In order to check different protein conformations, we tested different sized and cholesterol-content reconstituted apoAI particles (rHDL). Meanwhile cholesterol-free discs were active to induce mobilization, only small cholesterol-containing rHDL were active. To test the influence of a central domain in such events, we used two apoAI variants: one, with its central Y helix pair replaced by the C-terminal domain, and the other having a lysine deleted in central region. The helix-swapping variant decrease the cholesterol pool available to acyl-CoA cholesterol acyl transferase and increase mobilization of newly synthesized cholesterol. Instead, the deletion mutant had no effect on both events. We conclude that the central domain of apoAI is involved in cholesterol cell traffic and solubilization, and that a Y-type charge distribution in polar face may be required, as well as a correct helices-polar face orientation.
Gonzalez
Marina C
MC
Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP), CONICET/UNLP, Facultad de Ciencias Médicas, Calles 60 y 120, La Plata 1900, Buenos Aires, Argentina.
Toledo
Juan D
JD
Tricerri
M Alejandra
MA
Garda
Horacio A
HA
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D016632:Apolipoprotein A-I / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D016466:CHO Cells; D002784:Cholesterol / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D008076:Cholesterol, HDL / Q000737:chemistry / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D007424:Intracellular Fluid / Q000737:chemistry / Q000378:metabolism*; D008053:Lipid Mobilization / Q000235:genetics / Q000502:physiology*; D008239:Lysine / Q000235:genetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D017433:Protein Structure, Secondary / Q000235:genetics; D017434:Protein Structure, Tertiary; D012995:Solubility; D055672:Static Electricity
D016632:Apolipoprotein A-I; D008076:Cholesterol, HDL; D010455:Peptides; D002784:Cholesterol; D008239:Lysine