Isolation and Characterization of a New Member of the Insect Defensin Family from a Beetle, Allomyrina dichotoma

Miyanoshita, Akihiro; Hara, Seiichi; Sugiyama, Masao; Asaoka, Ai; Taniai, Kiyoko; Yukuhiro, Fumiko; Yamakawa, Minoru

doi:10.1006/bbrc.1996.0438

Cited by 69 publications

(59 citation statements)

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“…The first nine amino acids of the 19L-30R-NH # fragment are identical with the C-terminus of the presumed α-helical structure region, by sequence identity with some other insect defensins [7], suggesting that this fragment has the potential to form an α-helical structure. CD spectra of the 19L-30R-NH # fragment were measured using a spectropolarimeter in the absence or presence of PG\CL (3 : 1, mol\mol) liposomes.…”

Section: Spectramentioning

confidence: 85%

“…This peptide is composed of 43 amino acid residues, and has been shown to be effective against MRSA [7]. In the present study, we determined the active centre of defensin by measuring the anti-bacterial activity of 64 overlapping 12-mer peptides derived from it, and investigated the mode of action of this fragment.…”

Section: Introductionmentioning

confidence: 99%

“…Most anti-bacterial peptides have broadspectrum activities and disrupt bacterial membranes via a peptide-lipid interaction, unlike the mechanism of action of antibiotics used currently [5]. Some anti-bacterial peptides have been shown to be effective against antibiotic-resistant bacteria [6][7][8], suggesting their potential use as therapeutic agents. The cytotoxicity and antigenicity of these peptides, however, make them unsuitable for direct therapeutic use.…”

Section: Introductionmentioning

confidence: 99%

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Synthesis and characterization of bactericidal oligopeptides designed on the basis of an insect anti-bacterial peptide

Saido-Sakanaka

Ishibashi

Sagisaka

et al. 1999

Biochemical Journal

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Defensin from a beetle, Allomyrina dichotoma, is known to have anti-bacterial activity against Gram-positive bacteria. This peptide, which comprises 43 amino acid residues, was effective against methicillin-resistant Staphylococcus aureus. We identified the active site of beetle defensin by measuring anti-bacterial activity against S. aureus of 64 overlapping 12-mer peptides with either a free carboxylate or a free amide group at their C-termini. An LCAAHCLAIGRR-NH2 (19L–30R-NH2) fragment showed the greatest activity of the synthetic oligopeptides. The 19L–30R-NH2 fragment was effective against both Gram-positive and Gram-negative bacteria. CD spectra showed that the 19L–30R-NH2 fragment formed an α-helical structure in the lipidic environment. The anti-bacterial effect of the 19L–30R-NH2 fragment was due to its interaction with bacterial membranes, judging from the leakage of liposome-entrapped glucose. Its anti-bacterial activity was increased when certain amino acid residues were replaced. Truncated peptides having had some amino acids removed from the N-terminus of the 19L–30R-NH2 fragment (8–10-mer peptides) still had strong anti-bacterial activity. Deleting some amino acids from the C-terminal region of the fragment dramatically reduced activity, indicating that the C-terminal region of the 19L–30R-NH2 fragment, i.e. RR-NH2, is important for exerting anti-bacterial activity. The AHCLAIGRR-NH2 (22A–30R-NH2) fragment and its analogues exhibited about 3-fold and 9–12-fold higher activity against S. aureus than did the 19L–30R-NH2 fragment, and these analogues were effective against methicillin-resistant S. aureus and Pseudomonas aeruginosa isolated from patients. These oligopeptides showed no haemolytic activity and did not inhibit the growth of murine fibroblast cells.

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Section: Spectramentioning

confidence: 85%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Synthesis and characterization of bactericidal oligopeptides designed on the basis of an insect anti-bacterial peptide

Saido-Sakanaka

Ishibashi

Sagisaka

et al. 1999

Biochemical Journal

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“…5 Antimicrobial peptides are integral components of the innate immune system of all living organisms, including mammals, plants and insects. 6 Antimicrobial peptides result in strong natural defense in arthropod, particularly antimicrobial peptides of silk moth 7,8 and beetle, 9 which belong to the defensin family. Persulcatusin (IP), a tick antimicrobial peptide found in the midgut of Ixodes persulcatus, exhibits antimicrobial activity against Gram-positive bacteria such as S. aureus.…”

Section: Introductionmentioning

confidence: 99%

Activity of tick antimicrobial peptide from Ixodes persulcatus (persulcatusin) against cell membranes of drug-resistant Staphylococcus aureus

et al. 2016

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Persulcatusin (IP), which is an antimicrobial peptide found in Ixodes persulcatus midgut, is active against Gram-positive bacteria such as Staphylococcus aureus. Multidrug-resistant bacteria in particular methicillin-resistant S. aureus (MRSA), vancomycinintermediate S. aureus (VISA) and vancomycin-resistant S. aureus (VRSA) are a worldwide clinical concern. In the present study, to explore the potential of IP as a new agent against multidrug-resistant S. aureus infections, we evaluated the antimicrobial activity of IP against multidrug-resistant S. aureus strains by MIC 90 , morphological observation with scanning electron microscope (SEM), and the calcein leakage assay of membrane integrity. Among the six antimicrobial peptides used in this study, IP exhibited the lowest MIC 90 values for both vancomycin-susceptible and -resistant S. aureus strains. The IP MIC 90 against a VISA strain was equivalent to vancomycin, while the MIC 90 against VRSA was relatively low. SEM observations indicated that bacterial cells exposed to IP were crumpled and showed prominent structural changes. Moreover, IP influenced the cell membranes of both MRSA and VRSA in a mere 5 min, leading to leakage of the preloaded calcein. Although a VISA strain was resistant to the action of IP on cell membrane, the MIC 90 of IP was lower than that of Nisin, suggesting that IP had another bactericidal mechanism in addition to cell membrane attack. Our results indicate that the synthetic tick antimicrobial peptide, IP exhibits strong antibacterial activity against multidrug-resistant S. aureus strains, including VRSA, via both cell membrane attack and another unknown mechanism. IP represents a promising candidate for a new anti-VRSA therapy. The appearance of antibiotic-resistant bacteria has resulted in bacterial diseases re-emerging as a threat after the many decades since the introduction of the first antibiotic penicillin. 1 The development of new antibacterial agents is urgently required, and antimicrobial peptides are viewed as ideal candidate agents. 5 Antimicrobial peptides are integral components of the innate immune system of all living organisms, including mammals, plants and insects. 6 Antimicrobial peptides result in strong natural defense in arthropod, particularly antimicrobial peptides of silk moth 7,8 and beetle, 9 which belong to the defensin family. Persulcatusin (IP), a tick antimicrobial peptide found in the midgut of Ixodes persulcatus, exhibits antimicrobial activity against Gram-positive bacteria such as S. aureus. 10-12 Furthermore, we have reported previously that S. aureus strains could not be isolated from I. persulcatus during feeding. 13 This is attributable to the antimicrobial activity of IP, which is highly expressed during blood feeding. 10,11 We hypothesized that IP could exhibit antimicrobial activity against VISA and VRSA, as well as methicillin-susceptible S. aureus and MRSA.In this study, we evaluated the antimicrobial activity of IP against multidrug-resistant S. aureus strains using MIC, scanning electron m...

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“…Various papers have reported the isolation and description of anti-microbial defensins in different insects, but not in Lepidoptera (Chalk et al, 1995;Lowenberger et al, 1995;Cho et al, 1996Cho et al, , 1997Miyanoshita et al, 1996;Richman et al, 1996;Dimopoulos et al, 1997;Gao et al, 1999;Muller et al, 1999;Eggleston et al, 2000). The only data available on defensin in Lepidoptera concern Heliothis virescens, in which the inducible molecule, heliomicin, has been found.…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization of a defensin in the IZD‐MB‐0503 cell line derived from immunocytes of the insect Mamestra brassicae (Lepidoptera)

Mandrioli¹,

Bugli²,

Saltini³

et al. 2003

Biology of the Cell

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The induction of anti-microbial peptides against Gram positive and negative bacteria in the IZD-MB-0503 cell line from the lepidopteran Mamestra brassicae is demonstrated, while no anti-fungal activity is detected. The identification of a defensin-like molecule active against Gram positive bacteria is described for the first time in Lepidoptera. This molecule shows between 43% and 59% homology with group A defensins from other dipteran and hymenopteran species.

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Isolation and Characterization of a New Member of the Insect Defensin Family from a Beetle, Allomyrina dichotoma

Cited by 69 publications

References 0 publications

Synthesis and characterization of bactericidal oligopeptides designed on the basis of an insect anti-bacterial peptide

Synthesis and characterization of bactericidal oligopeptides designed on the basis of an insect anti-bacterial peptide

Activity of tick antimicrobial peptide from Ixodes persulcatus (persulcatusin) against cell membranes of drug-resistant Staphylococcus aureus

Molecular characterization of a defensin in the IZD‐MB‐0503 cell line derived from immunocytes of the insect Mamestra brassicae (Lepidoptera)

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