Solution and Solid-State Nuclear Magnetic Resonance Structural Investigations of the Antimicrobial Designer Peptide GL13K in Membranes

Nicole Harmouche; Christopher Aisenbrey; Fernando Porcelli; Youlin Xia; Sarah E Nelson; Xi Chen; Jesus Raya; Louic Vermeer; Conrado Aparicio; Gianluigi Veglia; Sven-Ulrik Gorr; Burkhard Bechinger

doi:10.1021/acs.biochem.7b00526

Solution and Solid-State Nuclear Magnetic Resonance Structural Investigations of the Antimicrobial Designer Peptide GL13K in Membranes

Nicole Harmouche, Christopher Aisenbrey, Fernando Porcelli, Youlin Xia, Sarah E Nelson, Xi Chen, Jesus Raya, Louic Vermeer, Conrado Aparicio, Gianluigi Veglia, Sven-Ulrik Gorr, Burkhard Bechinger

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

Abstract

The antimicrobial peptide GL13K encompasses 13 amino acid residues and has been designed and optimized from the salivary protein BPIFA2 to exhibit potent bacteriocidal and anti-biofilm activity against Gram-negative and Gram-positive bacteria as well as anti-lipopolysaccharide activity in vitro and in vivo. Here, the peptide was analyzed in a variety of membrane environments by circular dichroism spectroscopy and by high-resolution multidimensional solution nuclear magnetic resonance (NMR) spectroscopy. Whereas in the absence of membranes a random coil conformation predominates, the peptide adopts a helical structure from residue 5 to 11 in the presence of dodecylphosphocholine micelles. In contrast, a predominantly β-sheet structure was observed in the presence of lipid bilayers carrying negatively charged phospholipids. Whereas ¹⁵N solid-state NMR spectra are indicative of a partial alignment of the peptide ¹⁵N-¹H vector along the membrane surface, ²H and ³¹P solid-state NMR spectra indicate that in this configuration the peptide exhibits pronounced disordering activities on the phospholipid membrane, which is possibly related to antimicrobial action. GL13K, thus, undergoes a number of conformational transitions, including a random coil state in solution, a helical structure upon dilution at the surface of zwitterionic membranes, and β-sheet conformations at high peptide:lipid ratios.

Original language	English (US)
Pages (from-to)	4269-4278
Number of pages	10
Journal	Biochemistry
Volume	56
Issue number	32
DOIs	https://doi.org/10.1021/acs.biochem.7b00526
State	Published - Aug 15 2017

Bibliographical note

Funding Information:
This research was supported by Grant 1R01DE017989 from the National Institute for Dental and Craniofacial Research (S.-U.G.), Grant GM 64742 from the National Institute for General Medical Sciences (G.V.), and a fellowship supported by Grant R90DE023058 from the National Institute of Dental and Craniofacial Research (X.C.). Research funds were provided by the University of Minnesota School of Dentistry (S.-U.G.) and The Office of the Vice-president for Research at the University of Minnesota Project 55466 of the Grant-in-Aid of Research, Artistry and Scholarship Program (C. Aparicio). B.B. was supported by a sabbatical stay at the University of Minnesota School of Dentistry that was generously supported by the Schools Lasby Visiting Professor Fellowship and the University of Strasbourg. B.B. received additional support from the Agence Nationale de la Recherche (Projects TRANSPEP 07-PCV-0018 membraneDNP 12-BSV5-0012, and MemPepSyn 14-CE34-0001-01 and the LabEx Chemistry of Complex Systems 10-LABX-0026-CSC), the RTRA International Center of Frontier Research in Chemistry, the French Foundation for Medical Research (FRM), the University of Strasbourg, the CNRS, and the Region Alsace.

Publisher Copyright:
© 2017 American Chemical Society.

Publisher link

10.1021/acs.biochem.7b00526

Find It

Find It at U of M Libraries

Cite this

Harmouche, N., Aisenbrey, C., Porcelli, F., Xia, Y., Nelson, S. E., Chen, X., Raya, J., Vermeer, L., Aparicio, C., Veglia, G., Gorr, S-U., & Bechinger, B. (2017). Solution and Solid-State Nuclear Magnetic Resonance Structural Investigations of the Antimicrobial Designer Peptide GL13K in Membranes. Biochemistry, 56(32), 4269-4278. https://doi.org/10.1021/acs.biochem.7b00526

Harmouche, N, Aisenbrey, C, Porcelli, F, Xia, Y, Nelson, SE, Chen, X, Raya, J, Vermeer, L, Aparicio, C, Veglia, G , Gorr, S-U & Bechinger, B 2017, 'Solution and Solid-State Nuclear Magnetic Resonance Structural Investigations of the Antimicrobial Designer Peptide GL13K in Membranes', Biochemistry, vol. 56, no. 32, pp. 4269-4278. https://doi.org/10.1021/acs.biochem.7b00526

@article{6d23e0cecee24e7eb3b7f330a95bdf72,

title = "Solution and Solid-State Nuclear Magnetic Resonance Structural Investigations of the Antimicrobial Designer Peptide GL13K in Membranes",

abstract = "The antimicrobial peptide GL13K encompasses 13 amino acid residues and has been designed and optimized from the salivary protein BPIFA2 to exhibit potent bacteriocidal and anti-biofilm activity against Gram-negative and Gram-positive bacteria as well as anti-lipopolysaccharide activity in vitro and in vivo. Here, the peptide was analyzed in a variety of membrane environments by circular dichroism spectroscopy and by high-resolution multidimensional solution nuclear magnetic resonance (NMR) spectroscopy. Whereas in the absence of membranes a random coil conformation predominates, the peptide adopts a helical structure from residue 5 to 11 in the presence of dodecylphosphocholine micelles. In contrast, a predominantly β-sheet structure was observed in the presence of lipid bilayers carrying negatively charged phospholipids. Whereas 15N solid-state NMR spectra are indicative of a partial alignment of the peptide 15N-1H vector along the membrane surface, 2H and 31P solid-state NMR spectra indicate that in this configuration the peptide exhibits pronounced disordering activities on the phospholipid membrane, which is possibly related to antimicrobial action. GL13K, thus, undergoes a number of conformational transitions, including a random coil state in solution, a helical structure upon dilution at the surface of zwitterionic membranes, and β-sheet conformations at high peptide:lipid ratios.",

author = "Nicole Harmouche and Christopher Aisenbrey and Fernando Porcelli and Youlin Xia and Nelson, {Sarah E} and Xi Chen and Jesus Raya and Louic Vermeer and Conrado Aparicio and Gianluigi Veglia and Sven-Ulrik Gorr and Burkhard Bechinger",

note = "Funding Information: This research was supported by Grant 1R01DE017989 from the National Institute for Dental and Craniofacial Research (S.-U.G.), Grant GM 64742 from the National Institute for General Medical Sciences (G.V.), and a fellowship supported by Grant R90DE023058 from the National Institute of Dental and Craniofacial Research (X.C.). Research funds were provided by the University of Minnesota School of Dentistry (S.-U.G.) and The Office of the Vice-president for Research at the University of Minnesota Project 55466 of the Grant-in-Aid of Research, Artistry and Scholarship Program (C. Aparicio). B.B. was supported by a sabbatical stay at the University of Minnesota School of Dentistry that was generously supported by the Schools Lasby Visiting Professor Fellowship and the University of Strasbourg. B.B. received additional support from the Agence Nationale de la Recherche (Projects TRANSPEP 07-PCV-0018 membraneDNP 12-BSV5-0012, and MemPepSyn 14-CE34-0001-01 and the LabEx Chemistry of Complex Systems 10-LABX-0026-CSC), the RTRA International Center of Frontier Research in Chemistry, the French Foundation for Medical Research (FRM), the University of Strasbourg, the CNRS, and the Region Alsace. Publisher Copyright: {\textcopyright} 2017 American Chemical Society.",

year = "2017",

month = aug,

day = "15",

doi = "10.1021/acs.biochem.7b00526",

language = "English (US)",

volume = "56",

pages = "4269--4278",

journal = "Biochemistry",

issn = "0006-2960",

publisher = "American Chemical Society",

number = "32",

}

TY - JOUR

T1 - Solution and Solid-State Nuclear Magnetic Resonance Structural Investigations of the Antimicrobial Designer Peptide GL13K in Membranes

AU - Harmouche, Nicole

AU - Aisenbrey, Christopher

AU - Porcelli, Fernando

AU - Xia, Youlin

AU - Nelson, Sarah E

AU - Chen, Xi

AU - Raya, Jesus

AU - Vermeer, Louic

AU - Aparicio, Conrado

AU - Veglia, Gianluigi

AU - Gorr, Sven-Ulrik

AU - Bechinger, Burkhard

N1 - Funding Information: This research was supported by Grant 1R01DE017989 from the National Institute for Dental and Craniofacial Research (S.-U.G.), Grant GM 64742 from the National Institute for General Medical Sciences (G.V.), and a fellowship supported by Grant R90DE023058 from the National Institute of Dental and Craniofacial Research (X.C.). Research funds were provided by the University of Minnesota School of Dentistry (S.-U.G.) and The Office of the Vice-president for Research at the University of Minnesota Project 55466 of the Grant-in-Aid of Research, Artistry and Scholarship Program (C. Aparicio). B.B. was supported by a sabbatical stay at the University of Minnesota School of Dentistry that was generously supported by the Schools Lasby Visiting Professor Fellowship and the University of Strasbourg. B.B. received additional support from the Agence Nationale de la Recherche (Projects TRANSPEP 07-PCV-0018 membraneDNP 12-BSV5-0012, and MemPepSyn 14-CE34-0001-01 and the LabEx Chemistry of Complex Systems 10-LABX-0026-CSC), the RTRA International Center of Frontier Research in Chemistry, the French Foundation for Medical Research (FRM), the University of Strasbourg, the CNRS, and the Region Alsace. Publisher Copyright: © 2017 American Chemical Society.

PY - 2017/8/15

Y1 - 2017/8/15

N2 - The antimicrobial peptide GL13K encompasses 13 amino acid residues and has been designed and optimized from the salivary protein BPIFA2 to exhibit potent bacteriocidal and anti-biofilm activity against Gram-negative and Gram-positive bacteria as well as anti-lipopolysaccharide activity in vitro and in vivo. Here, the peptide was analyzed in a variety of membrane environments by circular dichroism spectroscopy and by high-resolution multidimensional solution nuclear magnetic resonance (NMR) spectroscopy. Whereas in the absence of membranes a random coil conformation predominates, the peptide adopts a helical structure from residue 5 to 11 in the presence of dodecylphosphocholine micelles. In contrast, a predominantly β-sheet structure was observed in the presence of lipid bilayers carrying negatively charged phospholipids. Whereas 15N solid-state NMR spectra are indicative of a partial alignment of the peptide 15N-1H vector along the membrane surface, 2H and 31P solid-state NMR spectra indicate that in this configuration the peptide exhibits pronounced disordering activities on the phospholipid membrane, which is possibly related to antimicrobial action. GL13K, thus, undergoes a number of conformational transitions, including a random coil state in solution, a helical structure upon dilution at the surface of zwitterionic membranes, and β-sheet conformations at high peptide:lipid ratios.

AB - The antimicrobial peptide GL13K encompasses 13 amino acid residues and has been designed and optimized from the salivary protein BPIFA2 to exhibit potent bacteriocidal and anti-biofilm activity against Gram-negative and Gram-positive bacteria as well as anti-lipopolysaccharide activity in vitro and in vivo. Here, the peptide was analyzed in a variety of membrane environments by circular dichroism spectroscopy and by high-resolution multidimensional solution nuclear magnetic resonance (NMR) spectroscopy. Whereas in the absence of membranes a random coil conformation predominates, the peptide adopts a helical structure from residue 5 to 11 in the presence of dodecylphosphocholine micelles. In contrast, a predominantly β-sheet structure was observed in the presence of lipid bilayers carrying negatively charged phospholipids. Whereas 15N solid-state NMR spectra are indicative of a partial alignment of the peptide 15N-1H vector along the membrane surface, 2H and 31P solid-state NMR spectra indicate that in this configuration the peptide exhibits pronounced disordering activities on the phospholipid membrane, which is possibly related to antimicrobial action. GL13K, thus, undergoes a number of conformational transitions, including a random coil state in solution, a helical structure upon dilution at the surface of zwitterionic membranes, and β-sheet conformations at high peptide:lipid ratios.

UR - http://www.scopus.com/inward/record.url?scp=85027419138&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85027419138&partnerID=8YFLogxK

U2 - 10.1021/acs.biochem.7b00526

DO - 10.1021/acs.biochem.7b00526

M3 - Article

C2 - 28699734

AN - SCOPUS:85027419138

SN - 0006-2960

VL - 56

SP - 4269

EP - 4278

JO - Biochemistry

JF - Biochemistry

IS - 32

ER -

Solution and Solid-State Nuclear Magnetic Resonance Structural Investigations of the Antimicrobial Designer Peptide GL13K in Membranes

Abstract

Bibliographical note

Publisher link

Find It

Other files and links

Fingerprint

Cite this