Membrane-dependent oligomeric structure and pore formation of a β-hairpin antimicrobial peptide in lipid bilayers from solid-state NMR

Mani, Rajeswari; Cady, Sarah D.; Tang, Ming; Waring, Alan J.; Lehrer, Robert I.; Hong, Mei

doi:10.1073/pnas.0605079103

Cited by 234 publications

(361 citation statements)

References 49 publications

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“…Each 19 F label existed in a two-spin cluster within the 15-Å upper limit of the technique. 77 Thus, the basic unit of PG-1 assembly is a NCCN dimer, which further associates into a . .…”

Section: Protegrin-1mentioning

confidence: 99%

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Structure and dynamics of cationic membrane peptides and proteins: Insights from solid‐state NMR

Hong¹,

Su²

2011

Protein Science

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130

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Many membrane peptides and protein domains contain functionally important cationic Arg and Lys residues, whose insertion into the hydrophobic interior of the lipid bilayer encounters significant energy barriers. To understand how these cationic molecules overcome the free energy barrier to insert into the lipid membrane, we have used solid-state NMR spectroscopy to determine the membrane-bound topology of these peptides. A versatile array of solid-state NMR experiments now readily yields the conformation, dynamics, orientation, depth of insertion, and site-specific protein-lipid interactions of these molecules. We summarize key findings of several Arg-rich membrane peptides, including b-sheet antimicrobial peptides, unstructured cell-penetrating peptides, and the voltage-sensing helix of voltage-gated potassium channels. Our results indicate the central role of guanidinium-phosphate and guanidinium-water interactions in dictating the structural topology of these cationic molecules in the lipid membrane, which in turn account for the mechanisms of this functionally diverse class of membrane peptides.

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“…Each 19 F label existed in a two-spin cluster within the 15-Å upper limit of the technique. 77 Thus, the basic unit of PG-1 assembly is a NCCN dimer, which further associates into a . .…”

Section: Protegrin-1mentioning

confidence: 99%

“…The peptide forms at least tetramers, which are no longer inserted into the membrane, as manifested by slow 1 H spin diffusion from the lipid chains. 77 Thus, PG-1 is prevented from inserting into the cholesterol-rich host cell membranes due to the membrane rigidity and negative curvature imparted by cholesterol.…”

Section: Protegrin-1mentioning

confidence: 99%

Structure and dynamics of cationic membrane peptides and proteins: Insights from solid‐state NMR

Hong¹,

Su²

2011

Protein Science

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130

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“…The proposed structure of the arenicin-2 tetramer is closely resembles the tetramerization motif of protegrin-1 revealed by solid-state NMR spectroscopy in anionic lipid membranes. 38 This b-hairpin peptide forms the tight dimers by parallel association of the C-terminal strands. These dimers, in turn, pack in higher-order aggregates by loose parallel association of the N-terminal strands with anionic lipid head groups, also possibly intercalated between them.…”

Section: Arenicin-2 Expression and Purificationmentioning

confidence: 99%

“…These dimers, in turn, pack in higher-order aggregates by loose parallel association of the N-terminal strands with anionic lipid head groups, also possibly intercalated between them. 38 Using all the available structural data on arenicin-2 (structure of the monomer in aqueous solution, 12 structure of the dimer and proposed model of the tetramer in anisotropic membrane mimetics) we can easily adapt the modern model describing mechanism of AMP membrane action (carpet/toroidal-pore model) [35][36][37] to the arenicin case ( Figure 9B). Initially, the peptide in the twisted b-hairpin conformation without pronounced amphipathicity approaches the anionic membrane surface ( Figure 9B).…”

Section: Arenicin-2 Expression and Purificationmentioning

confidence: 99%

Molecular insight into mechanism of antimicrobial action of the β‐hairpin peptide arenicin: Specific oligomerization in detergent micelles

et al. 2007

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Arenicins are 21‐residue cationic antimicrobial peptides isolated from marine polychaeta Arenicola marina. The peptides exhibit potent broad‐spectrum antimicrobial activity. In water solution arenicin‐2 adopts a β‐hairpin conformation, stabilized by one disulfide and nine hydrogen bonds. To determine the propensity for the peptide oligomerization in membrane mimetic systems, the recombinant arenicin‐2 was overexpressed as a fused form in Escherichia coli. The arenicin‐2 oligomerization and intermolecular packing in membrane mimicking environment were investigated using high‐resolution NMR spectroscopy. The present studies show that arenicin‐2 preserves a β‐hairpin structure and forms asymmetric dimers upon incorporation into the dodecylphosphocholine micelle. Two monomers of arenicin‐2 are aligned parallel to each other by the N‐terminal strands of the β‐hairpin (CN↑↑NC type of association). Polyacrylamide gel electrophoresis analysis indicated that in environment of anionic SDS micelles the arenicin‐2 might undergo further oligomerization and form tetramers. Our results afford further molecular insight into possible mechanism of antimicrobial action of arenicins. © 2007 Wiley Periodicals, Inc. Biopolymers 89: 455–464, 2008.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

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“…This method has been well-established for measuring the distances of proteins to the center of the lipid bilayer and to water. 40 The 1 H magnetization from mobile lipid chains and water was first selected by using a 1 H T 2 filter and was then transferred to the rigid oligocholates during a mixing period (t m ). A 1 H 180°pulse was applied in the middle of the T 2 filter to refocus the isotropic chemical shift.…”

Section: ■ Experimental Section Synthesis Of Compound 8 (Lt)mentioning

confidence: 99%

Aggregation and Dynamics of Oligocholate Transporters in Phospholipid Bilayers Revealed by Solid-State NMR Spectroscopy

et al. 2012

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Macrocycles made of cholate building blocks were previously found to transport glucose readily across lipid bilayers. In this study, an 15N, 13Cα-labeled glycine was inserted into a cyclic cholate trimer and attached at the end of a linear trimer, respectively. The isotopic labeling allowed us to use solid-state NMR spectroscopy to study the dynamics, aggregation, and depth of insertion of these compounds in lipid membranes. The cyclic compound was found to be mostly immobilized in DLPC, POPC/POPG, and POPC/POPG/cholesterol membranes, whereas the linear trimer displayed large-amplitude motion that depended on the membrane thickness and viscosity. 13C-detected 1H spin diffusion experiments revealed the depth of insertion of the compounds in the membranes, as well as their contact with water molecules. The data support a consistent stacking model for the cholate macrocycles in lipid membranes, driven by the hydrophobic interactions of the water molecules in the interior of the macrocycles. The study also shows a strong preference of the linear trimer for the membrane surface, consistent with its lack of transport activity in earlier liposome leakage assays. Disciplines Chemistry CommentsReprinted (adapted) with permission from Langmuir 28 (2012) ABSTRACT: Macrocycles made of cholate building blocks were previously found to transport glucose readily across lipid bilayers. In this study, an 15 N, 13 Cα-labeled glycine was inserted into a cyclic cholate trimer and attached at the end of a linear trimer, respectively. The isotopic labeling allowed us to use solid-state NMR spectroscopy to study the dynamics, aggregation, and depth of insertion of these compounds in lipid membranes. The cyclic compound was found to be mostly immobilized in DLPC, POPC/POPG, and POPC/POPG/cholesterol membranes, whereas the linear trimer displayed large-amplitude motion that depended on the membrane thickness and viscosity. 13 C-detected 1 H spin diffusion experiments revealed the depth of insertion of the compounds in the membranes, as well as their contact with water molecules. The data support a consistent stacking model for the cholate macrocycles in lipid membranes, driven by the hydrophobic interactions of the water molecules in the interior of the macrocycles. The study also shows a strong preference of the linear trimer for the membrane surface, consistent with its lack of transport activity in earlier liposome leakage assays.

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Membrane-dependent oligomeric structure and pore formation of a β-hairpin antimicrobial peptide in lipid bilayers from solid-state NMR

Cited by 234 publications

References 49 publications

Structure and dynamics of cationic membrane peptides and proteins: Insights from solid‐state NMR

Structure and dynamics of cationic membrane peptides and proteins: Insights from solid‐state NMR

Molecular insight into mechanism of antimicrobial action of the β‐hairpin peptide arenicin: Specific oligomerization in detergent micelles

Aggregation and Dynamics of Oligocholate Transporters in Phospholipid Bilayers Revealed by Solid-State NMR Spectroscopy

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