Melectin: A Novel Antimicrobial Peptide from the Venom of the Cleptoparasitic Bee <i>Melecta albifrons</i>

Čeřovský, Václav; Hovorka, Oldřich; Cvačka, Josef; Vobůrka, Zdeněk; Bednářová, Lucie; Borovičková, Lenka; Slaninová, Jiřina; Fučı́k, Vladimı́r

doi:10.1002/cbic.200800476

Cited by 55 publications

(35 citation statements)

References 39 publications

(47 reference statements)

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“…8). These studies have unveiled the distribution of amphiphilic and biologically active peptides such as melectin from Mellecta albifrons (Apoidea: Melectini) [24], codesane from Colletes daviesanus (Colletidae) [25], osmin from Osma rufa (Megachilidae) [26], lasioglossins from Lasioglossum laticeps (Halicitidae) [27], halictines from Halictus sexcinctus (Halicitidae) [28], macropin from Macropis fulvipes (Melittidae) [29] in the bee venom. It is interesting to note that the amino acid sequences of Xac-1 and Xac-2 are similar to those of melectin and osmin isolated from the long-tonged bees, but not to those of bombolitins and melittin isolated from the social bee venom, though carpenter bees, bumblebees and honeybees are closely related.…”

Section: Discussionmentioning

confidence: 99%

Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata

Kawakami

Goto

Murata

et al. 2017

J Venom Anim Toxins Incl Trop Dis

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BackgroundMass spectrometry-guided venom peptide profiling is a powerful tool to explore novel substances from venomous animals in a highly sensitive manner. In this study, this peptide profiling approach is successfully applied to explore the venom peptides of a Japanese solitary carpenter bee, Xylocopa appendiculata (Hymenoptera: Apoidea: Apidae: Anthophila: Xylocopinae: Xylocopini). Although interesting biological effects of the crude venom of carpenter bees have been reported, the structure and biological function of the venom peptides have not been elucidated yet.MethodsThe venom peptide profiling of the crude venom of X. appendiculata was performed by matrix-assisted laser desorption/ionization-time of flight mass spectroscopy. The venom was purified by a reverse-phase HPLC. The purified peptides were subjected to the Edman degradation, MS/MS analysis, and/or molecular cloning methods for peptide sequencing. Biological and functional characterization was performed by circular dichroism analysis, liposome leakage assay, and antimicrobial, histamine releasing and hemolytic activity tests.ResultsThree novel peptides with m/z 16508, 1939.3, and 1900.3 were isolated from the venom of X. appendiculata. The peptide with m/z 16508 was characterized as a secretory phospholipase A2 (PLA2) homolog in which the characteristic cysteine residues as well as the active site residues found in bee PLA2s are highly conserved. Two novel peptides with m/z 1939.3 and m/z 1900.3 were named as Xac-1 and Xac-2, respectively. These peptides are found to be amphiphilic and displayed antimicrobial and hemolytic activities. The potency was almost the same as that of mastoparan isolated from the wasp venom.ConclusionWe found three novel biologically active peptides in the venom of X. appendiculata and analyzed their molecular functions, and compared their sequential homology to discuss their molecular diversity. Highly sensitive mass analysis plays an important role in this study.Electronic supplementary materialThe online version of this article (doi:10.1186/s40409-017-0119-6) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata

Kawakami

Goto

Murata

et al. 2017

J Venom Anim Toxins Incl Trop Dis

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“…Xylopin has also a Pro10 residue in the sequence, in a similar position as both melectin and osmin that present a Pro11. According to Cerovsky et al [4], the Pro11 residue conferred to this peptide selectivity to the antimicrobial activity, as well as low hemolytic activity.…”

Section: Discussionmentioning

confidence: 99%

“…The first study on solitary bee venoms was published only in 2008 about the European solitary bee Melecta albifrons [4]. A novel peptide, melectin, was isolated and characterized.…”

Section: Introductionmentioning

confidence: 99%

Peptidomic analysis of the venom of the solitary bee Xylocopa appendiculata circumvolans

Kazuma

Ando

Nihei

et al. 2017

J Venom Anim Toxins Incl Trop Dis

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BackgroundAmong the hymenopteran insect venoms, those from social wasps and bees – such as honeybee, hornets and paper wasps – have been well documented. Their venoms are composed of a number of peptides and proteins and used for defending their nests and themselves from predators. In contrast, the venoms of solitary wasps and bees have not been the object of further research. In case of solitary bees, only major peptide components in a few venoms have been addressed. Therefore, the aim of the present study was to explore the peptide component profile of the venom from the solitary bee Xylocopa appendiculata circumvolans by peptidomic analysis with using LC-MS.MethodsA reverse-phase HPLC connected to ESI-OrbiTrap MS was used for LC-MS. On-line mass fingerprinting was made from TIC, and data-dependent tandem mass spectrometry gave MSMS spectra. A major peptide component was isolated by reverse-phase HPLC by conventional way, and its sequence was determined by Edman degradation, which was finally corroborated by solid phase synthesis. Using the synthetic specimen, biological activities (antimicrobial activity, mast cell devaluation, hemolysis, leishmanicidal activity) and pore formation in artificial lipid bilayer were evaluated.ResultsOn-line mass fingerprinting revealed that the crude venom contained 124 components. MS/MS analysis gave 75 full sequences of the peptide components. Most of these are related to the major and novel peptide, xylopin. Its sequence, GFVALLKKLPLILKHLH-NH2, has characteristic features of linear cationic α-helical peptides; rich in hydrophobic and basic amino acids with no disulfide bond, and accordingly, it can be predicted to adopt an amphipathic α-helix secondary structure. In biological evaluation, xylopin exhibited broad-spectrum antimicrobial activity, and moderate mast cell degranulation and leishmanicidal activities, but showed virtually no hemolytic activity. Additionally, the peptide was able to incorporate pores in artificial lipid bilayers of azolectin, confirming the mechanism of the cytolytic activity by pore formation in biological membranes.ConclusionsLC-ESI-MS and MS/MS analysis of the crude venom extract from a solitary bee Xylocopa appendiculata circumvolans revealed that the component profile of this venom mostly consisted of small peptides. The major peptide components, xylopin and xylopinin, were purified and characterized in a conventional manner. Their chemical and biological characteristics, belonging to linear cationic α-helical peptides, are similar to the known solitary bee venom peptides, melectin and osmin. Pore formation in artificial lipid bilayers was demonstrated for the first time with a solitary bee peptide.

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“…[34] The Petri dishes (90 mm in diameter) contained 20 mL of LB agar (Sigma). Melted "soft" agar (2 mL), prepared from LB broth (Sigma) and 0.5 % agar (Difco) containing about 10 7 colony-forming units of various bacteria (M. luteus, B. subtilis, S. aureus) was poured over the surface.…”

Section: Determination Of Antimicrobial Activitymentioning

confidence: 99%

Lucifensin, a Novel Insect Defensin of Medicinal Maggots: Synthesis and Structural Study

et al. 2011

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Recently, we identified a new insect defensin, named lucifensin that is secreted/excreted by the blowfly Lucilia sericata larvae into a wound as a disinfectant during the medicinal process known as maggot therapy. Here, we report the total chemical synthesis of this peptide of 40 amino acid residues and three intramolecular disulfide bridges by using three different protocols. Oxidative folding of linear peptide yielded a peptide with a pattern of disulfide bridges identical to that of native lucifensin. The synthetic lucifensin was active against Gram-positive bacteria and was not hemolytic. We synthesized three lucifensin analogues that are cyclized through one native disulfide bridge in different positions and having the remaining four cysteines substituted by alanine. Only the analogue cyclized through a Cys16-Cys36 disulfide bridge showed weak antimicrobial activity. Truncating lucifensin at the N-terminal by ten amino acid residues resulted in a drop in antimicrobial activity. Linear lucifensin having all six cysteine residues alkylated was inactive. Circular dichroism spectra measured in the presence of α-helix-promoting compounds showed different patterns for lucifensin and its analogues. Transmission electron microscopy revealed that Bacillus subtilis treatment with lucifensin induced significant changes in its envelope.

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Melectin: A Novel Antimicrobial Peptide from the Venom of the Cleptoparasitic Bee Melecta albifrons

Cited by 55 publications

References 39 publications

Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata

Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata

Peptidomic analysis of the venom of the solitary bee Xylocopa appendiculata circumvolans

Lucifensin, a Novel Insect Defensin of Medicinal Maggots: Synthesis and Structural Study

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