title: Formation of stable nanodiscs by bihelical apolipoprotein A-I mimetic peptide.
abstract: Nanodiscs are composed of scaffold protein or peptide such as apolipoprotein A-I (apoA-I) and phospholipids. Although peptide-based nanodiscs have an advantage to modulate the size of nanodiscs by changing phospholipid/peptide ratios, they are usually less stable than apoA-I-based nanodiscs. In this study, we designed a novel nanodisc scaffold peptide (NSP) that has proline-punctuated bihelical amphipathic structure based on apoA-I mimetic peptides. NSP formed α-helical structure on 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC) nanodiscs prepared by cholate dialysis method. Dynamic light scattering measurements demonstrated that diameters of NSP nanodiscs vary depending upon POPC/NSP ratios. Comparison of thermal unfolding of nanodiscs monitored by circular dichroism measurements demonstrated that NSP forms much more stable nanodiscs with POPC than monohelical peptide, 4F, exhibiting comparable stability to apoA-I-POPC nanodiscs. Intrinsic Trp fluorescence measurements showed that Trp residues of NSP exhibit more hydrophobic environment than that of 4 F on nanodiscs, suggesting the stronger interaction of NSP with phospholipids. Thus, the bihelical structure of NSP appears to increase the stability of nanodiscs because of the enhanced interaction of peptides with phospholipids. In addition, NSP as well as 4F spontaneously solubilized POPC vesicles into nanodiscs without using detergent. These results indicate that bihelical NSP forms nanodiscs with comparable stability to apoA-I and has an ability to control the size of nanodiscs simply by changing phospholipid/peptide ratios.
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kariyazono
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Nadai
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Rin
authors.initials: R
authors.lastname: Miyajima
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima, 770-8505, Japan.|Division of Drugs, National Institute of Health Sciences, Tokyo, 158-8501, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Takechi-Haraya
authors.affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, 305-8565, Japan.
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Baba
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Shigenaga
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Keiichiro
authors.initials: K
authors.lastname: Okuhira
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Otaka
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Saito
mesh_terms: D000595:Amino Acid Sequence; D016632:Apolipoprotein A-I / Q000737:chemistry*; D019826:Cholic Acid / Q000737:chemistry; D000067493:Dynamic Light Scattering; D006801:Humans; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D049329:Nanostructures / Q000737:chemistry*; D010316:Particle Size; D010446:Peptide Fragments / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D055550:Protein Stability; D017433:Protein Structure, Secondary
chemical_list: D016632:Apolipoprotein A-I; D010446:Peptide Fragments; D010713:Phosphatidylcholines; D019826:Cholic Acid; C028694:1-palmitoyl-2-oleoylphosphatidylcholine
keywords: apoA-I mimetic peptide; bihelical; nanodisc; nanodisc scaffold peptide