Lipid-Free Structure and Stability of Apolipoprotein A-I: Probing the Central Region by Mutation
| Autor: | Irina N. Gorshkova, David Atkinson, Vassilis I. Zannis, Tong Liu |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Protein Denaturation
Protein Folding Hot Temperature Apolipoprotein B medicine.disease_cause Biochemistry Central region Protein Structure Secondary medicine Humans Particle Size Guanidine Sequence Deletion Initial rate Mutation Liposome Apolipoprotein A-I biology Chemistry Cholesterol HDL Lipids Fluorescence Peptide Fragments Recombinant Proteins Receptor–ligand kinetics Kinetics Crystallography Spectrometry Fluorescence Amino Acid Substitution Liposomes Mutagenesis Site-Directed Phosphatidylcholines biology.protein lipids (amino acids peptides and proteins) Dimyristoylphosphatidylcholine Protein Binding Lipoprotein |
| Zdroj: | Biochemistry. 41:10529-10539 |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | To probe the structure and stability of the central region of lipid-free apolipoprotein (apo) A-I (residues 123-165), we studied the effects of four mutations made in this region on the conformation, stability, dimyristoylphosphatidylcholine (DMPC) binding kinetics, and size of discoidal reconstituted high-density lipoprotein (rHDL) particles. The apoA-I deletion delta(144-165) leads to a red shift in the wavelength of maximum fluorescence and a reduction in the alpha-helical content, the stability, the initial rate of association with DMPC liposomes, and the size of the discoidal particles. The data are consistent with the helical structure of residues 144-165, and the deletion appears to perturb the tertiary organization of the N-terminal half of apoA-I. In contrast, the deletion of the adjacent region, delta(136-143), leads to stabilization without altering the number of residues in the helical conformation or the initial rate of association with DMPC liposomes. The quadruple substitution E125K/E128K/K133E/E139K leads to approximately 17 additional residues in the helical conformation and an increase in the stability, the initial rate of association with DMPC liposomes, and the size of the rHDL particles. The findings are consistent with the disordered structure of the segment of residues 123-142, which becomes helical as a result of the quadruple mutation or upon lipid binding. The naturally occurring mutation L141R (also associated with coronary heart disease) that is located in this segment does not change the protein conformation but leads to a reduced stability and a decreased rate of association with DMPC liposomes that may relate to the observed altered functions of this mutant. |
| Databáze: | OpenAIRE |
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