title: Electrostatic effects on the stability of discoidal high-density lipoproteins.
abstract: High-density lipoproteins (HDL) remove cholesterol from peripheral tissues and thereby help to prevent atherosclerosis. Nascent HDL are discoidal complexes composed of a phospholipid bilayer surrounded by protein alpha-helices that are thought to form extensive stabilizing interhelical salt bridges. Earlier we showed that HDL stability, which is necessary for HDL functions, is modulated by kinetic barriers. Here we test the role of electrostatic interactions in the kinetic stability by analyzing the effects of salt, pH, and point mutations on model discoidal HDL reconstituted from human apolipoprotein C-1 (apoC-1) and dimyristoyl phosphatidylcholine (DMPC). Circular dichroism, Trp fluorescence, and light scattering data show that molar concentrations of NaCl or Na(2)SO(4) increase the apparent melting temperature of apoC-1:DMPC complexes by up to 20 degrees C and decelerate protein unfolding. Arrhenius analysis shows that 1 M NaCl stabilizes the disks by deltaDeltaG* approximately equal 3.5 kcal/mol at 37 degrees C and increases the activation energy of their denaturation and fusion by deltaE(a) approximately equal deltaDeltaH* approximately equal 13 kcal/mol, indicating that the salt-induced stabilization is enthalpy-driven. Denaturation studies in various solvent conditions (pH 5.7-8.2, 0-40% sucrose, 0-2 M trimethylamine N-oxide) suggest that the salt-induced disk stabilization results from ionic screening of unfavorable short-range Coulombic interactions. Thus, the dominant electrostatic interactions in apoC-1:DMPC disks are destabilizing. Comparison of the salt effects on the protein:lipid complexes of various composition reveals an inverse correlation between the lipoprotein stability and the salt-induced stabilization and suggests that short-range electrostatic interactions significantly contribute to lipoprotein stability: the better-optimized these interactions are, the more stable the complex is.
authors.affiliation: Department of Physiology and Biophysics, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Benjwal
authors.forename: Shobini
authors.initials: S
authors.lastname: Jayaraman
authors.forename: Olga
authors.initials: O
authors.lastname: Gursky
mesh_terms: D000595:Amino Acid Sequence; D016632:Apolipoprotein A-I / Q000737:chemistry; D053302:Apolipoprotein C-I; D001056:Apolipoproteins C / Q000737:chemistry; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D006801:Humans; D008027:Light; D008074:Lipoproteins / Q000737:chemistry*; D008075:Lipoproteins, HDL / Q000737:chemistry*; D008969:Molecular Sequence Data; D012542:Scattering, Radiation; D012965:Sodium Chloride; D055672:Static Electricity; D013431:Sulfates; D013816:Thermodynamics
chemical_list: D016632:Apolipoprotein A-I; D053302:Apolipoprotein C-I; D001056:Apolipoproteins C; D008074:Lipoproteins; D008075:Lipoproteins, HDL; D013431:Sulfates; C034366:discoidal lipoproteins; C012036:sodium sulfate; D012965:Sodium Chloride; D004134:Dimyristoylphosphatidylcholine