Creation of Cross-Linked Bilayer Membranes That Can Incorporate Membrane Proteins from Oligo-Asp-Based Peptide Gemini Surfactants

Koeda, Shuhei; Umezaki, Katsunari; Sumino, Ayumi; Noji, Tomoyasu; Ikeda, Atsushi; Yamämoto, Yasushi; Dewa, Takehisa; Taga, Keijiro; Nango, Mamoru; Tanaka, Toshiki; Mizuno, Toshihisa

doi:10.1021/la401566h

Cited by 7 publications

(6 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Owing to dimerization of single-chain surfactants, critical aggregation concentrations (CACs) of gemini surfactants are generally smaller than those of the parent single-chain surfactants . As for PG-surfactants, short linear peptides function not only in linking two long alkyl chains into a single molecule, but also as a hydrophilic headgroup. − To endow PG-surfactants with amphiphilic properties, the choice of hydrophilic linker peptides is indispensable. However, by adopting hydrophilic peptides at the peripheral peptide (Y-), less hydrophilic peptides can be chosen at linker peptides, without diminishing solubility in an aqueous buffer, enabling a flexible design of various functional PG-surfactants.…”

Section: Resultsmentioning

confidence: 99%

“…We recently developed a novel class of solubilization surfactants for membrane proteins based on 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 alkylamide methylene-modified Cys residues. , We named this scaffold the “peptide gemini surfactant” (PG-surfactant, Figure ) in our previous study . Choosing the betaine peptide sequence -Asp-Lys-Asp-Lys- as the linker peptide (-X-) and the N-terminal acetylated Asp or Lys as the peripheral peptide (Y-), we successfully constructed DKDKC 12 K and DKDKC 12 D as new solubilization surfactants for membrane proteins .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Design of New Extraction Surfactants for Membrane Proteins from Peptide Gemini Surfactants

et al. 2016

Self Cite

View full text Add to dashboard Cite

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, NPDGCKK, 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 NPDGCKK 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 NPDGCKK 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 NPDGCKK functions as a novel class of peptide-containing extraction surfactants for membrane proteins.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Design of New Extraction Surfactants for Membrane Proteins from Peptide Gemini Surfactants

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…Recently, we studied the evolution of several functional molecules from peptide-gemini surfactant (PG-surfactant) molecular framework. − The PG-surfactants (Figure ) are a kind of lipopeptide having a gemini-type molecular structure, consisting of two alkyl-chain-modified cysteine residues, a short linker peptide (-X-) between them, and periphery peptides at the N- and C-terminal sides (Y- and -Z, respectively) . In our previous study, by choosing suitable peptide sequences for -X-, Y-, and -Z, we succeeded in designing bilayer-forming amphiphiles, , that could act as solubilization surfactants , and extraction surfactants for membrane proteins; furthermore, these exhibited antimicrobial activities without hemolysis of blood erythrocytic cells . Interestingly, in spite of a simple basic molecular framework, we found a clear relationship between the peptide sequences at the X-, Y-, and Z-positions and the resulting properties and functions of the PG-surfactants.…”

Section: Introductionmentioning

confidence: 99%

“…16−21 The PG-surfactants (Figure 1) are a kind of lipopeptide having a gemini-type molecular structure, consisting of two alkyl-chain-modified cysteine residues, a short linker peptide (-X-) between them, and periphery peptides at the N-and C-terminal sides (Y-and -Z, respectively). 16 In our previous study, by choosing suitable peptide sequences for -X-, Y-, and -Z, we succeeded in designing bilayer-forming amphiphiles, 16,17 that could act as solubilization surfactants 18,19 and extraction surfactants 20 for membrane proteins; furthermore, these exhibited antimicrobial activities without hemolysis of blood erythrocytic cells. 21 Interestingly, in spite of a simple basic molecular framework, we found a clear relationship between the peptide sequences at the X-, Y-, and Z-positions and the resulting properties and functions of the PG-surfactants.…”

Section: ■ Introductionmentioning

confidence: 99%

Development of Cell-Penetration PG-Surfactants and Its Application in External Peptide Delivery to Cytosol

et al. 2020

Self Cite

View full text Add to dashboard Cite

Recently, development of techniques to deliver pharmacologically active biomacromolecules such as peptides and proteins to cytosol has gained much interest. Here, we applied the peptide gemini (PG)-surfactants to a novel platform to design cell penetration lipopeptides (CP-PGs), which can deliver exogenous peptides and proteins to cytosol. Among the number of candidate CP-PGs having different peptide sequences at the X-, Y-, and Z-positions, we focused on those having two C12 alkyl chains appended to the side chain of two Cys residues, the betaine sequence -Asp-Lys-Asp-Lys-between the alkylated Cys residues (i.e., at the X-position), and having different cationic peptide sequences of oligo-Lys or oligo-Arg at the Y-and/or Z-positions. With respect to cytotoxicity for mammalian cells such as NIH3T3 cells upon 1 h exposure, those having (Lys) 3 (K 3 -DKDKC 12 and DKCK 12 -K 3 ) showed lower cytotoxicity (IC 50 = 241 and 198 μM) among those having oligo-Lys, (Lys) n (n = 1, 3, 5; IC 50 = 88−197 μM). Similar lower cytotoxicity was also observed for the CP-PG having two (Lys) 3 at both N-and C-terminal sides (K 3 -DKDKC 12 -K 3 ) (IC 50 = 225 μM). In contrast, the CP-PG having (Arg) 3 at the N-terminal side (R 3 -DKDKC 12 ) showed higher cytotoxicity (IC 50 = 88 μM). Carrier abilities of the CP-PGs for exogenous peptides were evaluated using the proapoptotic domain (PAD) peptide, which induces apoptosis by disturbing mitochondrial membranes after delivery into cytosol. As a result, the CP-PGs of K 3 -DKDKC 12 , DKCK 12 -K 3 , K 3 -DKDKC 12 -K 3 , DKCK 12 -K 5 , and R 3 -DKDKC 12 exhibited micromolar range carrier ability (the necessary half concentration to induce cell death (EC 50 ) by delivering PAD peptide to cytosol was 10, 6.2, 8.5, 5.8, and 11.5 μM, respectively). Especially, the carrier abilities of DKCK 12 -K 3 and DKCK 12 -K 5 were superior to the well-established cell penetration Arg-rich R8 peptide (EC 50 = 6.8 μM). Together, our results indicate that the PG-surfactant molecular framework could be a potential new platform to design efficient cell penetration carrier materials.

show abstract

“…These PG-surfactants (Figure ) are a kind of lipopeptide having a gemini molecular structure, which consists of two alkyl-chain-modified cysteine residues, a short linker peptide (-X-) between them, and periphery peptides at the N- and C-terminal sides (Y- and -Z, respectively) . In our previous study, by choosing the suitable peptide sequences for -X-, Y-, and -Z, we succeeded in designing bilayer-forming amphiphiles, , that could act as solubilization surfactants , and extraction surfactants for membrane proteins. Interestingly, in spite of a simple basic molecular framework, we found that there is a clear relationship between the peptide sequences at the X-, Y-, and Z-positions and the resulting properties and functions of the PG-surfactant.…”

Section: Introductionmentioning

confidence: 99%

Development of New Antimicrobial Agents from Cationic PG-Surfactants Containing Oligo-Lys Peptides

et al. 2018

Self Cite

View full text Add to dashboard Cite

Peptide gemini-surfactant (PG-surfactant), a kind of lipopeptide, is composed of a short linker peptide (X) between two alkyl-chain-modified Cys residues and peripheral peptides at the N-terminal (Y) and the C-terminal (Z) sides, respectively, of the alkylated Cys residues. In this study, we developed and examined a series of PG-surfactants containing two C12 saturated alkanes and oligo-Lys, arranged at the X-, Y-, or Z-positions. To arrange oligo-Lys at the Y- or Z-positions, a repeat sequence of -Asp-Lys-Asp-Lys- was used at the X-position. All of the PG-surfactants exhibited high antimicrobial activity against both Gram-positive and -negative bacteria. In addition to high antimicrobial activity, a low hemolysis activity is prerequisite for efficient intravenous administration. Among the synthesized PG-surfactants, those having -(Lys)3- at the Y- or Z-positions, i.e. K 3 -DKDKC 12 and DKDKC 12 K 3 , showed reasonably low hemolytic activities. This combination of high antimicrobial activity along with low hemolytic activity is an essential and unique property and has not been previously reported for the synthesized lipopeptides. Further, using scanning electron microscopy (SEM) and N-phenyl-1-naphthylamine (NPN) uptake assay we showed that the antimicrobial activity of these PG-surfactants may be attributed to membrane disruptive mechanisms. Although the PG-surfactants with low hemolytic activity could interact and localize onto red blood cell surfaces and cause slight expansion of cell morphologies, no subsequent penetration occurred. In summary, we describe here the successful development of PG-surfactants having high antibacterial and low hemolytic activity, thus providing a significant molecular platform to develop novel antimicrobial agents.

show abstract

Creation of Cross-Linked Bilayer Membranes That Can Incorporate Membrane Proteins from Oligo-Asp-Based Peptide Gemini Surfactants

Cited by 7 publications

References 27 publications

Design of New Extraction Surfactants for Membrane Proteins from Peptide Gemini Surfactants

Design of New Extraction Surfactants for Membrane Proteins from Peptide Gemini Surfactants

Development of Cell-Penetration PG-Surfactants and Its Application in External Peptide Delivery to Cytosol

Development of New Antimicrobial Agents from Cationic PG-Surfactants Containing Oligo-Lys Peptides

Contact Info

Product

Resources

About