title: Apolipoprotein A-I helical structure and stability in discoidal high-density lipoprotein (HDL) particles by hydrogen exchange and mass spectrometry.
abstract: To understand high-density lipoprotein (HDL) structure at the molecular level, the location and stability of α-helical segments in human apolipoprotein (apo) A-I in large (9.6 nm) and small (7.8 nm) discoidal HDL particles were determined by hydrogen-deuterium exchange (HX) and mass spectrometry methods. The measured HX kinetics of some 100 apoA-I peptides specify, at close to amino acid resolution, the structural condition of segments throughout the protein sequence and changes in structure and stability that occur on incorporation into lipoprotein particles. When incorporated into the large HDL particle, the nonhelical regions in lipid-free apoA-I (residues 45-53, 66-69, 116-146, and 179-236) change conformation from random coil to α-helix so that nearly the entire apoA-I molecule adopts helical structure (except for the terminal residues 1-6 and 237-243). The amphipathic α-helices have relatively low stability, in the range 3-5 kcal/mol, indicating high flexibility and dynamic unfolding and refolding in seconds or less. A segment encompassed by residues 125-158 exhibits bimodal HX labeling indicating co-existing helical and disordered loop conformations that interchange on a time scale of minutes. When incorporated around the edge of the smaller HDL particle, the increase in packing density of the two apoA-I molecules forces about 20% more residues out of direct contact with the phospholipid molecules to form disordered loops, and these are the same segments that form loops in the lipid-free state. The region of disc-associated apoA-I that binds the lecithin-cholesterol acyltransferase enzyme is well structured and not a protruding unstructured loop as reported by others.
authors: Lipid Research Group, Gastroenterology, Hepatology and Nutrition Division, Children's Hospital of Philadelphia, Philadelphia, PA 19104-4318, USA.
authors: Palaniappan
authors: P
authors: Sevugan Chetty
authors: Leland
authors: L
authors: Mayne
authors: Zhong-Yuan
authors: ZY
authors: Kan
authors: Sissel
authors: S
authors: Lund-Katz
authors: S Walter
authors: SW
authors: Englander
authors: Michael C
authors: MC
authors: Phillips
mesh_terms: D016632:Apolipoprotein A-I / Q000737:chemistry*; D041961:Deuterium Exchange Measurement; D006801:Humans; D008075:Lipoproteins, HDL / Q000737:chemistry*; D013058:Mass Spectrometry; D055550:Protein Stability; D017433:Protein Structure, Secondary
chemical_list: D016632:Apolipoprotein A-I; D008075:Lipoproteins, HDL