title: Free cholesterol determines reassembled high-density lipoprotein phospholipid phase structure and stability.
abstract: Reassembled high-density lipoproteins (rHDL) of various sizes and compositions containing apo A-I or apo A-II as their sole protein, dimyristoylphosphatidylcholine (DMPC), and various amounts of free cholesterol (FC) have been isolated and analyzed by differential scanning calorimetry (DSC) and by circular dichroism to determine their stability and the temperature dependence of their helical content. Our data show that the multiple rHDL species obtained at each FC mole percent usually do not have the same FC mole percent as the starting mixture and that the size of the multiple species increases in a quantized way with their respective FC mole percent. DSC studies reveal multiple phases or domains that can be classified as virtual DMPC, which contains a small amount of DMPC that slightly reduces the melting temperature (Tm), a boundary phase that is adjacent to the apo A-I or apo A-II that circumscribes the discoidal rHDL, and a mixed FC/DMPC phase that has a Tm that increases with FC mole percent. Only the large rHDL contain virtual DMPC, whereas all contain boundary phase and various amounts of the mixed FC/DMPC phase according to increasing size and FC mole percent. As reported by others, FC stabilizes the rHDL. For rHDL (apo A-II) compared to rHDL (apo A-I), this occurs in spite of the reduced number of helical regions that mediate binding to the DMPC surface. This effect is attributed to the very high lipophilicity of apo A-II and the reduction in the polarity of the interface between DMPC and the aqueous phase with an increasing FC mole percent, an effect that is expected to increase the strength of the hydrophobic associations with the nonpolar face of the amphipathic helices of apo A-II. These data are relevant to the differential effects of FC and apolipoprotein species on intracellular and plasma membrane nascent HDL assembly and subsequent remodeling by plasma proteins.
authors.affiliation: Cardiovascular and Thrombosis Research Section, Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, United States.
authors.forename: Matthew
authors.initials: M
authors.lastname: Auton
authors.forename: G Randall
authors.initials: GR
authors.lastname: Bassett
authors.forename: Baiba K
authors.initials: BK
authors.lastname: Gillard
authors.forename: Henry J
authors.initials: HJ
authors.lastname: Pownall
mesh_terms: D016632:Apolipoprotein A-I / Q000737:chemistry; D016633:Apolipoprotein A-II / Q000737:chemistry; D002152:Calorimetry, Differential Scanning; D008076:Cholesterol, HDL / Q000737:chemistry*; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry*; D006801:Humans; D044367:Phase Transition; D010743:Phospholipids / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D013816:Thermodynamics
chemical_list: D016632:Apolipoprotein A-I; D016633:Apolipoprotein A-II; D008076:Cholesterol, HDL; D010743:Phospholipids; D004134:Dimyristoylphosphatidylcholine