Bioactive peptides; Cryptides; Peptide diversity; Peptidomics; Wasp venom; Wasp Venoms; Peptides; Angiotensin-Converting Enzyme Inhibitors; Insect Proteins; Animals; Proteolysis; Amino Acid Sequence; Angiotensin-Converting Enzyme Inhibitors/chemistry; Species Specificity; Wasp Venoms/chemistry; Wasp Venoms/metabolism; Peptides/chemistry; Peptides/analysis; Peptides/metabolism; Proteomics/methods; Wasps; Biophysics; Biochemistry
Abstract :
[en] Wasp venoms possess complex compositions and diverse bioactivities, making them potential pharmacological sources. In this study, venoms from four wasp species (Vespa mandarinia, V. velutina, V. basalis, and Provespa barthelemyi) were collected by electrical stimulation and analyzed using liquid chromatography-tandem mass spectrometry. A total of 681 peptides were identified, nearly 90% of which had not been previously reported. Comparative analyses revealed pronounced species-specific signatures at both peptide and peptide family levels. Sequence-based analyses indicated that peptide release is consistent with targeted proteolytic cleavage patterns, exhibiting features resembling known substrate preferences of metalloproteases and serine proteases, rather than stochastic degradation. Bioinformatic predictions identified 291 peptides with potential bioactive properties spanning multiple functional categories, with angiotensin-converting enzyme (ACE) and dipeptidyl peptidase IV (DPP4) inhibitory activities being the most prominently represented. Among these, eight ACE inhibitory peptides and seventeen DPP4 inhibitory peptides were prioritized as candidates based on predicted safety profiles, and sequence-based analysis further identified ten putative cryptides. Overall, this study establishes the first comparative peptidomic dataset across four wasp venoms, providing insights into peptide diversity, inferred generation patterns, and predicted activities. SIGNIFICANCE: Venoms are rich sources of biologically active molecules and have historically provided templates for clinically used therapeutics. Although major protein toxins have been extensively characterized, the endogenous low-molecular-weight peptide fraction remains comparatively underexplored, particularly in social wasps, and systematic comparative resources remain limited. Venom peptidomic datasets inherently contain multiple layers of biological information, including diversification patterns, peptide origin, and potential bioactivity, yet these aspects are often interpreted independently. By integrating these dimensions, this study establishes a multi-level analytical framework for extracting biological insights and function-related information from venom peptidomes. The identification of consistent cleavage patterns suggests a degree of regulation in peptide generation, shifting the interpretation of venom peptides from degradation by-products toward biologically organized repertoires. Moreover, candidate prioritization illustrates how peptidomic datasets can generate experimentally testable functional hypotheses rather than serving solely as descriptive catalogs. The resulting dataset serves as a reference resource for cumulative comparative analyses. The analytical framework presented here also provides a transferable strategy for functional peptide discovery in other complex secretions.
Disciplines :
Physical, chemical, mathematical & earth Sciences: Multidisciplinary, general & others
Author, co-author :
Wang, Kai ; Université de Liège - ULiège > TERRA Research Centre > Entomologie, Phytopathologie et Productions Innovantes (EPPI)
Qiao, Jiangtao; School of Life Sciences and Food Engineering, Hebei University of Engineering, Handan 056038, China
Quinton, Loïc ; Université de Liège - ULiège > Département de chimie (sciences) > Chimie biologique
Eppe, Gauthier ; Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Meng, Xianbin; Equipment R&D Center, Shenzhen Medical Academy of Research and Translation, Shenzhen 518107, Guangdong, China
Huang, Guozhong; Longling Huangshi Bee Industry Co., Ltd, Longling County, 678300, Yunnan, China
Haubruge, Eric ; Université de Liège - ULiège > Département GxABT > Entomologie, Phytopathologie et Productions Innovantes (EPPI)
Zhang, Hongcheng; State Key Laboratory of Resource Insects, Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Beijing 100093, China. Electronic address: 460414874@qq.com
Language :
English
Title :
Comparative peptidomics of four wasp venoms reveals extensive peptide diversity, proteolytic patterns, and predicted bioactivities.
Modica, M.V., Sunagar, K., Holford, M., Dutertre, S., Diversity and evolution of animal venoms: neglected targets, ecological interactions, future perspectives. Front. Ecol. Evol., 8, 2020, 65, 10.3389/fevo.2020.00065.
Schendel, V., Rash, L.D., Jenner, R.A., Undheim, E.A., The diversity of venom: the importance of behavior and venom system morphology in understanding its ecology and evolution. Toxins, 11(11), 2019, 666, 10.3390/toxins11110666.
Freuville, L., Matthys, C., Quinton, L., Gillet, J.-P., Venom-derived peptides for breaking through the glass ceiling of drug development. Front. Chem., 12, 2024, 1465459, 10.3389/fchem.2024.1465459.
Messadi, E., Snake venom components as therapeutic drugs in ischemic heart disease. Biomolecules, 13(10), 2023, 1539, 10.3390/biom13101539.
Safavi-Hemami, H., Brogan, S.E., Olivera, B.M., Pain therapeutics from cone snail venoms: from Ziconotide to novel non-opioid pathways. J. Proteome 190 (2019), 12–20, 10.1016/j.jprot.2018.05.009.
Zhang, Z.-Q., Animal Biodiversity: An Outline of Higher-Level Classification and Survey of Taxonomic Richness. 2011, Magnolia press, Auckland, New Zealand.
Czaikoski, P.G., Menaldo, D.L., Marcussi, S., Baseggio, A.L., Fuly, A.L., Paula, R.C., Quadros, A.U., Romao, P.R., Buschini, M.L., Cunha, F.Q., Anticoagulant and fibrinogenolytic properties of the venom of Polybia occidentalis social wasp. Blood Coagul. Fibrinolysis 21:7 (2010), 653–659, 10.1097/MBC.0b013e32833cea7a.
Isidorov, V., Zalewski, A., Zambrowski, G., Swiecicka, I., Chemical composition and antimicrobial properties of honey bee venom. Molecules, 28(10), 2023, 4135, 10.3390/molecules28104135.
El-Seedi, H.R., Eid, N., Abd El-Wahed, A.A., Rateb, M.E., Afifi, H.S., Algethami, A.F., Zhao, C., Al Naggar, Y., Alsharif, S.M., Tahir, H.E., Honey bee products: preclinical and clinical studies of their anti-inflammatory and immunomodulatory properties. Front. Nutr., 8, 2022, 761267, 10.3389/fnut.2021.761267.
Kachel, H.S., Patel, R.N., Franzyk, H., Mellor, I.R., Block of nicotinic acetylcholine receptors by philanthotoxins is strongly dependent on their subunit composition. Sci. Rep., 6(1), 2016, 38116, 10.1038/srep38116.
Hanafi, M.Y., Zaher, E.L., El-Adely, S.E., Sakr, A., Ghobashi, A.H., Hemly, M.H., Kazem, A.H., Kamel, M.A., The therapeutic effects of bee venom on some metabolic and antioxidant parameters associated with HFD-induced non-alcoholic fatty liver in rats. Exp. Ther. Med. 15:6 (2018), 5091–5099, 10.3892/etm.2018.6028.
Jang, S., Kim, K.H., Clinical effectiveness and adverse events of bee venom therapy: a systematic review of randomized controlled trials. Toxins, 12(9), 2020, 558, 10.3390/toxins12090558.
Abd El-Wahed, A., Yosri, N., Sakr, H.H., Du, M., Algethami, A.F., Zhao, C., Abdelazeem, A.H., Tahir, H.E., Masry, S.H., Abdel-Daim, M.M., Wasp venom biochemical components and their potential in biological applications and nanotechnological interventions. Toxins, 13(3), 2021, 206, 10.3390/toxins13030206.
Sharma, S., Upadhyay, R.K., Hymenoptera toxins: biological activity, pharmaceutical and therapeutic uses. Int J Pharm Pharm Sci, 13, 2021 10.22159/ijpps.2021v13i4.40473.
Hemajha, L., Singh, S., Biji, C.A., Balde, A., Benjakul, S., Nazeer, R.A., A review on inflammation modulating venom proteins/peptide therapeutics and their delivery strategies: a review. Int. Immunopharmacol., 142, 2024, 113130, 10.1016/j.intimp.2024.113130.
Dallas, D.C., Guerrero, A., Parker, E.A., Robinson, R.C., Gan, J., German, J.B., Barile, D., Lebrilla, C.B., Current peptidomics: applications, purification, identification, quantification, and functional analysis. Proteomics 15:5–6 (2015), 1026–1038, 10.1002/pmic.201400310.
Clynen, E., Baggerman, G., Husson, S., Landuyt, B., Schoofs, L., Peptidomics in drug research. Expert Opin. Drug Discov. 3:4 (2008), 425–440, 10.1517/17460441.3.4.425.
Ito, L.T., Miyamoto, J.G., Sant'Anna, S.S., Grego, K.F., Tanaka-Azevedo, A.M., Tashima, A.K., Unveiling the peptidome diversity of Lachesis muta snake venom: discovery of novel fragments of metalloproteinase, L-amino acid oxidase, and bradykinin potentiating peptides. Biochem. Biophys. Res. Commun., 683, 2023, 149090, 10.1016/j.bbrc.2023.10.022.
Zeng, L., Zhang, C., Yang, M., Sun, J., Lu, J., Zhang, H., Qin, J., Zhang, W., Jiang, Z., Unveiling the diversity and modifications of short peptides in Buthus martensii scorpion venom through liquid chromatography-high resolution mass spectrometry. Toxins, 16(3), 2024, 155, 10.3390/toxins16030155.
Cologna, C.T., Rodrigues, R.S., Santos, J., De Pauw, E., Arantes, E.C., Quinton, L., Peptidomic investigation of Neoponera villosa venom by high-resolution mass spectrometry: seasonal and nesting habitat variations. J. Venom. Anim. Toxins Incl. Trop. Dis., 24(1), 2018, 6, 10.1186/s40409-018-0141-3.
Portes-Junior, J.A., Yamanouye, N., Carneiro, S.M., Knittel, P.S., Sant'Anna, S.v.S., Nogueira, F.C., Junqueira, M., Magalhães, G.S., Domont, G.B., Moura-da-Silva, A.M., Unraveling the processing and activation of snake venom metalloproteinases. J. Proteome Res. 13:7 (2014), 3338–3348 https://doi.org/10.1021/pr500185a.
Nicolau, C.A., Carvalho, P.C., Junqueira-de-Azevedo, I.L., Teixeira-Ferreira, A., Junqueira, M., Perales, J., Neves-Ferreira, A.G.C., Valente, R.H., An in-depth snake venom proteopeptidome characterization: benchmarking Bothrops jararaca. J. Proteome 151 (2017), 214–231 https://doi.org/10.1016/j.jprot.2016.06.029.
Tashima, A.K., Zelanis, A., Kitano, E.S., Ianzer, D., Melo, R.L., Rioli, V., Sant'anna, S.S., Schenberg, A.C., Camargo, A.C., Serrano, S.M., Peptidomics of three Bothrops snake venoms: insights into the molecular diversification of proteomes and peptidomes. Mol. Cell. Proteomics 11:11 (2012), 1245–1262, 10.1074/mcp.M112.019331.
Miyamoto, J.G., Kitano, E.S., Zelanis, A., Nachtigall, P.G., Junqueira-de-Azevedo, I., Sant'Anna, S.S., da Silva, R.L., Bersanetti, P.A., Carmona, A.K., Pereira, P.J.B., A novel metalloproteinase-derived cryptide from Bothrops cotiara venom inhibits angiotensin-converting enzyme activity. Biochimie 216 (2024), 90–98, 10.1016/j.biochi.2023.10.010.
Lex, A., Gehlenborg, N., Strobelt, H., Vuillemot, R., Pfister, H., UpSet: visualization of intersecting sets. IEEE Trans. Vis. Comput. Graph. 20:12 (2014), 1983–1992, 10.1109/TVCG.2014.2346248.
Minkiewicz, P., Iwaniak, A., Darewicz, M., BIOPEP-UWM database of bioactive peptides: current opportunities. Int. J. Mol. Sci., 20(23), 2019, 5978, 10.3390/ijms20235978.
Ueki, N., Someya, K., Matsuo, Y., Wakamatsu, K., Mukai, H., Cryptides: functional cryptic peptides hidden in protein structures. Pept. Sci. 88:2 (2007), 190–198, 10.1002/bip.20687.
Mukai, H., Seki, T., Nakano, H., Hokari, Y., Takao, T., Kawanami, M., Tsukagoshi, H., Kimura, H., Kiso, Y., Shimonishi, Y., Mitocryptide-2: purification, identification, and characterization of a novel cryptide that activates neutrophils. J. Immunol. 182:8 (2009), 5072–5080, 10.4049/jimmunol.0802965.
Perez-Riverol, A., Lasa, A.M., dos Santos-Pinto, J.R.A., Palma, M.S., Insect venom phospholipases A1 and A2: roles in the envenoming process and allergy. Insect Biochem. Mol. Biol. 105 (2019), 10–24, 10.1016/j.ibmb.2018.12.011.
Sukprasert, S., Rungsa, P., Uawonggul, N., Incamnoi, P., Thammasirirak, S., Daduang, J., Daduang, S., Purification and structural characterisation of phospholipase A1 (Vespapase, Ves a 1) from Thai banded tiger wasp (Vespa affinis) venom. Toxicon 61 (2013), 151–164 https://doi.org/10.1016/j.toxicon.2012.10.024.
Rungsa, P., Peigneur, S., Daduang, S., Tytgat, J., Purification and biochemical characterization of VesT1s, a novel phospholipase A1 isoform isolated from the venom of the greater banded wasp Vespa tropica. Toxicon 148 (2018), 74–84, 10.1016/j.toxicon.2018.03.015.
Yang, H., Xu, X., Ma, D., Zhang, K., Lai, R., A phospholipase A1 platelet activator from the wasp venom of Vespa magnifica (smith). Toxicon 51:2 (2008), 289–296, 10.1016/j.toxicon.2007.10.003.
Luo, L., Kamau, P.M., Lai, R., Bioactive peptides and proteins from wasp venoms. Biomolecules, 12(4), 2022, 527, 10.3390/biom12040527.
Barassé, V., Téné, N., Klopp, C., Paquet, F., Tysklind, N., Troispoux, V., Lalägue, H., Orivel, J., Lefranc, B., Leprince, J., Venomics survey of six myrmicine ants provides insights into the molecular and structural diversity of their peptide toxins. Insect Biochem. Mol. Biol., 151, 2022, 103876, 10.1016/j.ibmb.2022.103876.
Duran, L.H., Rymer, T.L., Wilson, D.T., Variation in venom composition in the Australian funnel-web spiders Hadronyche valida. Toxicon X, 8, 2020, 100063, 10.1016/j.toxcx.2020.100063.
Oldrati, V., Koua, D., Allard, P.-M., Hulo, N., Arrell, M., Nentwig, W., Lisacek, F., Wolfender, J.-L., Kuhn-Nentwig, L., Stöcklin, R., Peptidomic and transcriptomic profiling of four distinct spider venoms. PLoS One, 12(3), 2017, e0172966, 10.1371/journal.pone.0172966.
Leme, A.F.P., Escalante, T., Pereira, J.G., Oliveira, A.K., Sanchez, E.F., Gutiérrez, J.M., Serrano, S.M., Fox, J.W., High resolution analysis of snake venom metalloproteinase (SVMP) peptide bond cleavage specificity using proteome based peptide libraries and mass spectrometry. J. Proteome 74:4 (2011), 401–410, 10.1016/j.jprot.2010.12.002.
Bertholim, L., Zelanis, A., Oliveira, A.K., Serrano, S.M., Proteome-derived peptide library for the elucidation of the cleavage specificity of HF3, a snake venom metalloproteinase. Amino Acids 48:5 (2016), 1331–1335, 10.1007/s00726-016-2218-z.
Trevisan-Silva, D., Bednaski, A.V., Gremski, L.H., Chaim, O.M., Veiga, S.S., Senff-Ribeiro, A., Differential metalloprotease content and activity of three Loxosceles spider venoms revealed using two-dimensional electrophoresis approaches. Toxicon 76 (2013), 11–22, 10.1016/j.toxicon.2013.08.065.
Bordon, K.C., Santos, G.C., Martins, J.G., Wiezel, G.A., Amorim, F.G., Crasset, T., Redureau, D., Quinton, L., Procópio, R.E., Arantes, E.C., Pioneering comparative proteomic and enzymatic profiling of Amazonian scorpion venoms enables the isolation of their first α-Ktx, metalloprotease, and phospholipase A2. Toxins, 17(8), 2025, 411, 10.3390/toxins17080411.
Moreau, S.J., Asgari, S., Venom proteins from parasitoid wasps and their biological functions. Toxins 7:7 (2015), 2385–2412, 10.3390/toxins7072385.
Zelanis, A., Huesgen, P.F., Oliveira, A.K., Tashima, A.K., Serrano, S.M., Overall, C.M., Snake venom serine proteinases specificity mapping by proteomic identification of cleavage sites. J. Proteome 113 (2015), 260–267, 10.1016/j.jprot.2014.10.002.
Ullah, A., Masood, R., Ali, I., Ullah, K., Ali, H., Akbar, H., Betzel, C., Thrombin-like enzymes from snake venom: structural characterization and mechanism of action. Int. J. Biol. Macromol. 114 (2018), 788–811, 10.1016/j.ijbiomac.2018.03.164.
Choo, Y.M., Lee, K.S., Yoon, H.J., Kim, B.Y., Sohn, M.R., Roh, J.Y., Je, Y.H., Kim, N.J., Kim, I., Woo, S.D., Dual function of a bee venom serine protease: prophenoloxidase-activating factor in arthropods and fibrin (ogen) olytic enzyme in mammals. PLoS One, 5(5), 2010, e10393, 10.1371/journal.pone.0010393.
Becchimanzi, A., Avolio, M., Bostan, H., Colantuono, C., Cozzolino, F., Mancini, D., Chiusano, M.L., Pucci, P., Caccia, S., Pennacchio, F., Venomics of the ectoparasitoid wasp Bracon nigricans. BMC Genomics, 21(1), 2020, 34, 10.1186/s12864-019-6396-4.
Kalogeropoulos, K., Treschow, A.F., auf dem Keller, U., Escalante, T., Rucavado, A., Gutiérrez, J.M., Laustsen, A.H., Workman, C.T., Protease activity profiling of snake venoms using high-throughput peptide screening. Toxins, 11(3), 2019, 170, 10.3390/toxins11030170.
Moura-da-Silva, A.M., Almeida, M.T., Portes-Junior, J.A., Nicolau, C.A., Gomes-Neto, F., Valente, R.H., Processing of snake venom metalloproteinases: generation of toxin diversity and enzyme inactivation. Toxins, 8(6), 2016, 183, 10.3390/toxins8060183.
Dashevsky, D., Baumann, K., Undheim, E.A., Nouwens, A., Ikonomopoulou, M.P., Schmidt, J.O., Ge, L., Kwok, H.F., Rodriguez, J., Fry, B.G., Functional and proteomic insights into aculeata venoms. Toxins, 15(3), 2023, 224, 10.3390/toxins15030224.
Gatti, J.-L., Belghazi, M., Legeai, F., Ravallec, M., Frayssinet, M., Robin, S., Aboubakar-Souna, D., Srinivasan, R., Tamo, M., Poirié, M., Proteo-trancriptomic analyses reveal a large expansion of metalloprotease-like proteins in atypical venom vesicles of the wasp Meteorus pulchricornis (Braconidae). Toxins, 13(7), 2021, 502, 10.3390/toxins13070502.
dos Santos, L.D., Santos, K.S., Pinto, J.R.A., Dias, N.B., Souza, B.M.d., dos Santos, M.F., Perales, J., Domont, G.B., Castro, F.M., Kalil, J.E., Profiling the proteome of the venom from the social wasp Polybia paulista: a clue to understand the envenoming mechanism. J. Proteome Res. 9:8 (2010), 3867–3877, 10.1021/pr1000829.
Abd El-Hakam, F.E.-Z., Abo Laban, G., Badr El-Din, S., Abd El-Hamid, H., Farouk, M.H., Apitherapy combination improvement of blood pressure, cardiovascular protection, and antioxidant and anti-inflammatory responses in dexamethasone model hypertensive rats. Sci. Rep., 12(1), 2022, 20765, 10.1038/s41598-022-24727-z.
Hassan, A.K., El-kotby, D.A., Tawfik, M.M., Badr, R.E., Bahgat, I.M., Antidiabetic effect of the Egyptian honey bee (Apis mellifera) venom in alloxan-induced diabetic rats. J. Basic Appl. Zool., 80(1), 2019, 58, 10.1186/s41936-019-0127-x.
Rocha-Resende, C., Leão, N.M., de Lima, M.E., Santos, R.A., de Castro Pimenta, A.M., Verano-Braga, T., Moving pieces in a cryptomic puzzle: Cryptide from Tityus serrulatus Ts3 Nav toxin as potential agonist of muscarinic receptors. Peptides 98 (2017), 70–77, 10.1016/j.peptides.2016.12.018.
Guo, R., Guo, G., Wang, A., Xu, G., Lai, R., Jin, H., Spider-venom peptides: structure, bioactivity, strategy, and research applications. Molecules, 29(1), 2023, 35, 10.3390/molecules29010035.
