Crystallization and preliminary X-ray diffraction analysis of apolipoprotein E-containing lipoprotein particles

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Nov 1;61(Pt 11):981-4. doi: 10.1107/S1744309105032410. Epub 2005 Oct 20.

Abstract

High-resolution structural information is available for several soluble plasma apolipoproteins (apos) in a lipid-free state. However, this information provides limited insight into structure-function relationships, as this class of proteins primarily performs its functions of lipid transport and modulation of lipid metabolism in a lipid-bound state on lipoprotein particles. Here, the possibility of generating homogeneous lipoprotein particles that could be crystallized was explored, opening the possibility of obtaining high-resolution structural information by X-ray crystallography. To test this possibility, apoE4 complexed with the phospholipid dipalmitoylphosphatidylcholine was chosen. Uniform particles containing 50% lipid and 50% apoE4 were obtained and crystallized using the hanging-drop method. Two crystal forms diffract to beyond 8 A resolution.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 1,2-Dipalmitoylphosphatidylcholine / chemistry
  • Apolipoproteins E / chemistry*
  • Cross-Linking Reagents / pharmacology
  • Crystallization / methods
  • Crystallography, X-Ray
  • Electrophoresis, Polyacrylamide Gel
  • Fibroblasts / metabolism
  • Humans
  • Hydrogen-Ion Concentration
  • Lipids / chemistry
  • Lipoproteins / chemistry*
  • Microscopy, Electron
  • Phospholipids / chemistry
  • Protein Binding
  • Protein Conformation
  • Receptors, LDL / chemistry
  • Temperature
  • X-Ray Diffraction / methods

Substances

  • Apolipoproteins E
  • Cross-Linking Reagents
  • Lipids
  • Lipoproteins
  • Phospholipids
  • Receptors, LDL
  • 1,2-Dipalmitoylphosphatidylcholine