Stable isotope-assisted NMR characterization of interaction between lipid A and sarcotoxin IA, a cecropin-type antibacterial peptide

Yagi-Utsumi, Maho; Yamaguchi, Yoshiki; Boonsri, Pornthip; Iguchi, Takeshi; Okemoto, Kazuo; Natori, Shunji; Kato, Koichi

doi:10.1016/j.bbrc.2013.01.009

Cited by 9 publications

(6 citation statements)

References 25 publications

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“…In case of sarcotoxin IA, a 39-residue cecropin-type peptide, the peptide forms a helixhinge-helix structure in methanol solution similar to other cecropin-type peptides [46]. On the other hand, in a DPC/lipid A mixture, the peptide exhibits an α-helix at its Leu3-Arg18 sequence within the N-terminal region, whereas the C-terminal region, which forms an α-helix in methanol, was unstructured [45]. These chemical shift changes between samples with or without the DPC/lipid A mixture indicated that the charged residues Lys4 and Lys5 were involved in the interaction with lipid A.…”

Section: Discussionmentioning

confidence: 95%

“…Structure in LPS Critical residues for interaction with LPS Cecropin P1 SWLSKTAKKLENSAKKRISEGIAIAIQGGPR α-Helix in C-terminal (K15-G29) K15, K16, I22, I26 Sarcotoxin IA [44] GWLKKIGKKIERVGQHTRDATIQGLGIAQQAANVAATAR α-Helix in N-terminal (L3-R18) W2, K4, K5 Pa4 [45] GFFALIPKIISSPLFKTLLSAVGSALSSSGGQE Helix (L5-S12)-turn-helix (K16-G31) K8, K16, I9, I10, P13, L18, L19, A21…”

Section: Peptide [Reference] Sequencementioning

confidence: 99%

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Lipopolysaccharide-bound structure of the antimicrobial peptide cecropin P1 determined by nuclear magnetic resonance spectroscopy

et al. 2016

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Antimicrobial peptides (AMPs) are components of the innate immune system and may be potential alternatives to conventional antibiotics because they exhibit broad-spectrum antimicrobial activity. The AMP cecropin P1 (CP1), isolated from nematodes found in the stomachs of pigs, is known to exhibit antimicrobial activity against Gram-negative bacteria. In this study, we investigated the interaction between CP1 and lipopolysaccharide (LPS), which is the main component of the outer membrane of Gram-negative bacteria, using circular dichroism (CD) and nuclear magnetic resonance (NMR). CD results showed that CP1 formed an α-helical structure in a solution containing LPS. For NMR experiments, we expressed 15 N-and 13 C-labeled CP1 in bacterial cells and successfully assigned almost all backbone and side-chain proton resonance peaks of CP1 in water for transferred nuclear Overhauser effect (Tr-NOE) experiments in LPS. We performed 15 N-edited and 13 C-edited Tr-NOE spectroscopy (Tr-NOESY) for CP1 bound to LPS. Tr-NOE peaks were observed at the only Cterminal region of CP1 in LPS. The results of structure calculation indicated that the Cterminal region (Lys15-Gly29) formed the well-defined α-helical structure in LPS.Finally, the docking study revealed that Lys15/Lys16 interacted with phosphate at GlcN I via an electrostatic interaction and that Ile22/Ile26 was in close proximity with the acyl chain of lipid A.

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Section: Discussionmentioning

confidence: 95%

Section: Peptide [Reference] Sequencementioning

confidence: 99%

Lipopolysaccharide-bound structure of the antimicrobial peptide cecropin P1 determined by nuclear magnetic resonance spectroscopy

et al. 2016

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“…The two N-terminal residues (Gly 1 -Trp 2 ) of sarcotoxin-IA are important for the activity against E. coli since the two residues are required for binding of sarcotoxin-IA to the lipid A of LPS (Okemoto et al, 2002). Two N-terminal residues (Trp 2 and Phe 5 ) of papiliocin and H. cecropia cecropin-A are also important for interaction of cecropins with negatively charged bacterial cell membrane (Lee et al, 2013a), and N-terminal Lys 4 and Lys 5 of sarcotoxin-IA are key residues for the interaction with lipid A and antimicrobial activity (Yagi-Utsumi et al, 2013). In addition to antimicrobial activity, cecropins and cecropin derivatives (SB-37 and Shiva) are also active against parasites, including Plasmodium and Trypanosome (Arrowood et al, 1991; Barr et al, 1995; Boisbouvier et al, 1998; Boulanger et al, 2002a; Gwadz et al, 1989; Jaynes et al, 1988; Rodriguez et al, 1995), and can inhibit replication of HIV-1 virus (Wachinger et al, 1998) and proliferation of cancer cells (Chen et al, 1997; Suttmann et al, 2008).…”

Section: Cecropinsmentioning

confidence: 99%

“…H. cecropia cecropin-A (37 residues) contains two helical regions in residues 5–21 and 24–37 (Holak et al, 1988), and sarcotoxin-IA (39 residues) exhibits an N-terminal amphipathic α-helix (residues 3–23) and a more hydrophobic C-terminal α-helix (residues 28–38) connected by a hinge region (residues 24–27) (Iwai et al, 1993). The N-terminal amphiphilic α-helix (residues 3–18) of sarcotoxin-IA is formed upon interaction with micelles, and amino acids in this α-helix are involved in specific interaction with lipid A (Yagi-Utsumi et al, 2013). Papiliocin (37 residues) adopts an unordered structure in aqueous solution but converts to α-helical structure in the presence of SDS (sodium dodecyl sulfate), DPC (dodecylphosphocholine), HFIP (1,1,1,3,3,3-hexafluoro-2-propanol) and LPS micelles, with two amphipathic α-helixes (residues 3–21 and 25–36) linked by a hinge (Ala 22 -Gly 23 -Pro 24 ) (Fig.…”

Section: Cecropinsmentioning

confidence: 99%

Insect antimicrobial peptides and their applications

Huiuk

Chowdhury

Huang

et al. 2014

Appl Microbiol Biotechnol

483

459

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Insects are one of the major sources of antimicrobial peptides/proteins (AMPs). Since observation of antimicrobial activity in the hemolymph of pupae from the giant silk moths Samia Cynthia and Hyalophora cecropia in 1974 and purification of first insect AMP (cecropin) from H. cecropia pupae in 1980, over 150 insect AMPs have been purified or identified. Most insect AMPs are small and cationic, and they show activities against bacteria and/or fungi, as well as some parasites and viruses. Insect AMPs can be classified into four families based on their structures or unique sequences: the α-helical peptides (cecropin and moricin), cysteine-rich peptides (insect defensin and drosomycin), proline-rich peptides (apidaecin, drosocin and lebocin), and glycine-rich peptides/proteins (attacin and gloverin). Among insect AMPs, defensins, cecropins, proline-rich peptides and attacins are common, while gloverins and moricins have been identified only in Lepidoptera. Most active AMPs are small peptides of 20–50 residues, which are generated from larger inactive precursor proteins or pro-proteins, but gloverins (~14 kDa) and attacins (~20 kDa) are large antimicrobial proteins. In this mini-review, we will discuss current knowledge and recent progress in several classes of insect AMPs, including insect defensins, cecropins, attacins, lebocins and other proline-rich peptides, gloverins, and moricins, with a focus on structural-functional relationships and their potential applications.

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“…Growing evidence indicates that the aggregation of various amyloidogenic proteins such as Aβ and αSN can be highly enhanced at glycolipid membranes, suggesting that dynamical glycolipid-dependent conformational changes in proteins are a crucial step for the subsequent amyloid fibril formation that may lead to neurodegenerative disease. [16][17][18][19][20] However, no detailed structural study has reported on how proteins bind to glycolipid membranes. Conformational analysis of protein-glycolipid complexes is one of the most challenging subjects for NMR spectroscopy, because such complexes are relatively huge and unstable, with high fluidity and flexibility.…”

Section: Protein Molecular Assemblies and Their Pathological Functionsmentioning

confidence: 99%

NMR Characterization of Conformational Dynamics and Molecular Assemblies of Proteins

Yagi-Utsumi

2019

Biological & Pharmaceutical Bulletin

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Dynamic conformational transitions and molecular assemblies are essential properties of proteins, and relevant to their biological and pathological functions. Neurodegenerative diseases are known to be caused by abnormal, toxic assemblies of related proteins, e.g., amyloid β (Aβ) in Alzheimer's disease. Growing evidence indicates that the aggregation of various amyloidogenic proteins, including Aβ, can be highly enhanced at glycolipid membranes, suggesting that dynamic glycolipid-dependent conformational changes of proteins constitute crucial steps for their subsequent pathogenic amyloid fibril formation. It has also been proposed that several proteins, including molecular chaperones, can capture amyloidogenic proteins and thereby suppress their fibrillization. NMR spectroscopy provides a powerful tool for characterizing the conformational dynamics and intermolecular interactions of proteins, as well as for exploring transiently formed weak interactions among proteins in solution with various biomolecules, such as glycolipids. Our research group therefore attempted to elucidate the structural basis of protein-glycolipid and protein-protein interactions that either promote or suppress molecular assemblies of amyloidogenic proteins, using both solution and solid-state NMR methods in conjunction with other biophysical techniques. Our findings provide structural views of molecular processes involving amyloidogenic proteins of clinical and pathological interest and offer clues for the development of drugs to prevent and treat neurodegenerative diseases.

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Stable isotope-assisted NMR characterization of interaction between lipid A and sarcotoxin IA, a cecropin-type antibacterial peptide

Cited by 9 publications

References 25 publications

Lipopolysaccharide-bound structure of the antimicrobial peptide cecropin P1 determined by nuclear magnetic resonance spectroscopy

Lipopolysaccharide-bound structure of the antimicrobial peptide cecropin P1 determined by nuclear magnetic resonance spectroscopy

Insect antimicrobial peptides and their applications

NMR Characterization of Conformational Dynamics and Molecular Assemblies of Proteins

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