The carboxyl-terminal segment of apolipoprotein A-V undergoes a lipid-induced conformational change.
The carboxyl-terminal segment of apolipoprotein A-V undergoes a lipid-induced conformational change.
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DOI：
10.1021/bi1005859
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发表时间：
2010-06-15
期刊：
影响因子：
2.9
通讯作者：
Ryan, Robert O.
中科院分区：
文献类型：
--
作者：
Mauldin, Kasuen;Lee, Brian L.;Oleszczuk, Marta;Sykes, Brian D.;Ryan, Robert O.
Apolipoprotein (apo) A-V is a 343 residue, multi-domain protein that plays an important role in regulation of plasma triglyceride homeostasis. Primary sequence analysis revealed a unique tetra-proline sequence (Pro293 – Pro296) near the carboxyl terminus of the protein. A peptide corresponding to the 48 residue segment beyond the tetra-proline motif was generated from a recombinant apoA-V precursor wherein Pro295 was replaced by Met. Cyanogen bromide cleavage of the precursor protein, followed by negative affinity chromatography, yielded a purified peptide. Nondenaturing polyacrylamide gel electrophoresis verified that apoA-V(296-343) solubilizes phospholipid vesicles, forming a relatively heterogeneous population of reconstituted high density lipoprotein with Stoke's diameters > 17 nm. At the same time, apoA-V(296-343) failed to bind a spherical lipoprotein substrate in vitro. Far UV circular dichroism spectroscopy revealed the peptide is unstructured in buffer yet adopts significant α-helical secondary structure in the presence of the lipid mimetic solvent, trifluoroethanol (TFE; 50% v/v). Heteronuclear multidemensional NMR spectroscopy experiments were conducted with uniformly 15N and 15N/13C labeled peptide in 50 % TFE. Peptide backbone assignment and secondary structure prediction using TALOS+ reveals the peptide adopts α-helix secondary structure from residues 309 – 334. In TFE, apoA-V(296-343) adopts an extended amphipathic α-helix, consistent with a role in lipoprotein binding as a component of full-length apoA-V.
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