A novel amphipathic cell-penetrating peptide based on the N-terminal glycosaminoglycan binding region of human apolipoprotein E

Publication date: Available online 15 December 2018Source: Biochimica et Biophysica Acta (BBA) - BiomembranesAuthor(s): Takashi Ohgita, Yuki Takechi-Haraya, Ryo Nadai, Mana Kotani, Yuki Tamura, Karin Nishikiori, Kazuchika Nishitsuji, Kenji Uchimura, Koki Hasegawa, Kumiko Sakai-Kato, Kenichi Akaji, Hiroyuki SaitoAbstractIn the direct cell membrane penetration, arginine-rich cell-penetrating peptides are thought to penetrate into cells across the hydrophobic lipid membranes. To investigate the effect of the amphipathic property of arginine-rich peptide on the cell-penetrating ability, we designed a novel amphipathic cell-penetrating peptide, A2-17, and its derivative, A2-17KR, in which all lysine residues are substituted with arginine residues, based on the glycosaminoglycan binding region in the N-terminal α-helix bundle of human apolipoprotein E. Isothermal titration calorimetry showed that A2-17 variants have a strong ability to bind to heparin with high affinity. Circular dichroism and tryptophan fluorescence measurements demonstrated that A2-17 variants bind to lipid vesicles with a structural change from random coil to amphipathic α-helix, being inserted into the hydrophobic membrane interiors. Flow cytometric analysis and confocal laser scanning microscopy demonstrated the great cell penetration efficiency of A2-17 variants into CHO-K1 cells when incubated at low peptide concentrations (2 μM or less), suggesting that the increased amphipathicity with α-helix format...

Source: Biochimica et Biophysica Acta (BBA) Biomembranes - December 16, 2018 Category: Biochemistry Source Type: research