title: Design of New Extraction Surfactants for Membrane Proteins from Peptide Gemini Surfactants.
abstract: The development of additional extraction surfactants for membrane proteins is necessary for membrane protein research, since optimal combinations for the successful extraction of target membrane proteins from biological membranes that minimize protein denaturation are hard to predict. In particular, those that have a unique basal molecular framework are quite attractive and highly desired in this research field. In this study, we successfully constructed a new extraction surfactant for membrane proteins, NPDGC12KK, from the peptide-gemini-surfactant (PG-surfactant) molecular framework. The PG-surfactant is a U-shaped lipopeptide scaffold, consisting of a short linker peptide (-X-) between two long alkyl-chain-modified Cys residues and a peripheral peptide (Y-) at the N-terminal side of long alkyl-chain-modified Cys residues. Using photosystem I (PSI) and photosystem II (PSII) derived from Thermosynecoccus vulcanus as representative membrane proteins, we evaluated whether NPDGC12KK could solubilize membrane proteins while maintaining structure and functions. Neither the membrane integral domain nor the cytoplasmic domain of PSI and PSII suffered any damage upon the use of NPDGC12KK based on detailed photophysical measurements. Using thylakoid membranes of T. vulcanus as a representative biological membrane sample, we performed experiments to extract membrane proteins, such as PSI and PSII. Based on the extraction efficiency and maintenance of protein supramolecular structure established using clear native-PAGE analyses, we proved that NPDGC12KK functions as a novel class of peptide-containing extraction surfactants for membrane proteins.
authors: Graduate School of Engineering, Nagoya Institute of Technology , Gokiso-cho, Showa-ku, Nagoya, Aichi 466-8555, Japan.
authors: Masahide
authors: M
authors: Shibata
authors: Graduate School of Engineering, Nagoya Institute of Technology , Gokiso-cho, Showa-ku, Nagoya, Aichi 466-8555, Japan.
authors: Shuhei
authors: S
authors: Koeda
authors: Osaka City University , The OCU Advanced Research Institute for Natural Science & Technology (OCARINA), 3-3-138 Sugimoto-cho, Sumiyoshi, Osaka 558-8585, Japan.
authors: Tomoyasu
authors: T
authors: Noji
authors: Osaka City University , The OCU Advanced Research Institute for Natural Science & Technology (OCARINA), 3-3-138 Sugimoto-cho, Sumiyoshi, Osaka 558-8585, Japan.
authors: Keisuke
authors: K
authors: Kawakami
authors: Graduate School of Engineering, Nagoya Institute of Technology , Gokiso-cho, Showa-ku, Nagoya, Aichi 466-8555, Japan.
authors: Yuya
authors: Y
authors: Ido
authors: Graduate School of Pharmaceutical Sciences, Nagoya City University , 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan.
authors: Yuichi
authors: Y
authors: Amano
authors: Graduate School of Pharmaceutical Sciences, Nagoya City University , 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan.
authors: Naoki
authors: N
authors: Umezawa
authors: Graduate School of Pharmaceutical Sciences, Nagoya City University , 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan.
authors: Tsunehiko
authors: T
authors: Higuchi
authors: Graduate School of Engineering, Nagoya Institute of Technology , Gokiso-cho, Showa-ku, Nagoya, Aichi 466-8555, Japan.
authors: Takehisa
authors: T
authors: Dewa
authors: Graduate School of Science, Nagoya University , Furo-cho, Chikusa-ku Nagoya, Aichi 464-8602, Japan.
authors: Shigeru
authors: S
authors: Itoh
authors: Osaka City University , The OCU Advanced Research Institute for Natural Science & Technology (OCARINA), 3-3-138 Sugimoto-cho, Sumiyoshi, Osaka 558-8585, Japan.
authors: Nobuo
authors: N
authors: Kamiya
authors: Graduate School of Engineering, Nagoya Institute of Technology , Gokiso-cho, Showa-ku, Nagoya, Aichi 466-8555, Japan.
authors: Toshihisa
authors: T
authors: Mizuno
mesh_terms: D005591:Chemical Fractionation / Q000379:methods; D003545:Cysteine / Q000737:chemistry; D055666:Lipopeptides / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D008823:Micelles; D010455:Peptides / Q000737:chemistry; D045331:Photosystem I Protein Complex / Q000737:chemistry; D045332:Photosystem II Protein Complex / Q000737:chemistry; D015202:Protein Engineering / Q000379:methods; D013050:Spectrometry, Fluorescence; D013501:Surface-Active Agents / Q000737:chemistry*; D046939:Synechocystis / Q000737:chemistry; D020524:Thylakoids / Q000737:chemistry
chemical_list: D055666:Lipopeptides; D008565:Membrane Proteins; D008823:Micelles; D010455:Peptides; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D013501:Surface-Active Agents; D003545:Cysteine