Crystal Structure of Δ(185–243)apoA-I Suggests a Mechanistic Framework for the Protein Adaptation to the Changing Lipid Load in Good Cholesterol: From Flatland to Sphereland via Double Belt Belt-Buckle Double Hairpin and Trefoil/Tetrafoil

摘要

ApoA-I is the major protein of plasma high-density lipoproteins (HDL), macromolecular assemblies of proteins and lipids that remove cell cholesterol and protect against atherosclerosis. HDL heterogeneity, large size (7.7–12 nm) and ability to exchange proteins have prevented high-resolution structural analysis. Low-resolution studies showed that two apoA-I molecules form an antiparallel α-helical “double belt” around an HDL particle. Atomic-resolution structure of the C-terminal truncated lipid-free Δ(185–243)apoA-I, determined recently by Mei and Atkinson, provides unprecedented new insights into HDL structure-function. It allows us to propose a molecular mechanism for the adaptation of the full-length protein to increasing lipid load during cholesterol transport. ApoA-I conformations on the small, mid-size and large HDL are proposed based on the tandem α-helical repeats and the crystal structure of Δ(185–243)apoA-I, and are validated by comparison with extensive biophysical data reported by many groups. In our models, the central half of the double belt (“constant” segment 66–184) is structurally conserved while the N- and C-terminal half (“variable” segments 1–65 and 185–243) re-arranges upon HDL growth. This includes incremental unhinging of the N-terminal bundle around two flexible regions containing G39 and G65 to elongate the belt, along with concerted swing motion of the double belt around G65-P66 and G185–G186 hinges that are aligned on various-size particles, to confer 2D surface curvature to spherical HDL. The proposed conformational ensemble integrates and improves several existing HDL models. It helps provide a structural framework necessary to understand functional interactions with over 60 other HDL-associated proteins and, ultimately, improve cardioprotective function of HDL.