Helical structure and stability in human apolipoprotein A-I by hydrogen exchange and mass spectrometry.
Apolipoprotein A-I (apoA-I) stabilizes anti-atherogenic high density lipoprotein particles (HDL) in the circulation and governs their biogenesis, metabolism, and functional interactions. To decipher these important structure-function relationships, it will be necessary to understand the structure, stability, and plasticity of the apoA-I molecule. Biophysical studies show that lipid-free apoA-I contains a large amount of alpha-helical structure but the location of this structure and its properties are not established. We used hydrogen-deuterium exchange coupled with a fragmentation-separation method and mass spectrometric analysis to study human lipid-free apoA-I in its physiologically pertinent monomeric form. The acquisition of approximately 100 overlapping peptide fragments that redundantly cover the 243-residue apoA-I polypeptide made it possible to define the positions and stabilities of helical segments and to draw inferences about their interactions and dynamic properties. Residues 7-44, 54-65, 70-78, 81-115, and 147-178 form alpha-helices, accounting for a helical content of 48 +/- 3%, in agreement with circular dichroism measurements (49%). At 3 to 5 kcal/mol in free energy of stabilization, the helices are far more stable than could be achieved in isolation, indicating mutually stabilizing helix bundle interactions. However the helical structure is dynamic, unfolding and refolding in seconds, allowing facile apoA-I reorganization during HDL particle formation and remodeling.
Lipid Research Group, Gastroenterology, Hepatology and Nutrition Division, Children's Hospital of Philadelphia, Philadelphia, PA 19104-4318, USA.
Palaniappan Sevugan
PS
Chetty
Leland
L
Mayne
Sissel
S
Lund-Katz
David
D
Stranz
S Walter
SW
Englander
Michael C
MC
Phillips
D016632:Apolipoprotein A-I / Q000737:chemistry* / Q000378:metabolism*; D041961:Deuterium Exchange Measurement; D006801:Humans; D008075:Lipoproteins, HDL / Q000378:metabolism; D013058:Mass Spectrometry; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship
D016632:Apolipoprotein A-I; D008075:Lipoproteins, HDL