Assessing the Proteomic Activity of the Venom of the Ant Ectatomma tuberculatum (Hymenoptera: Formicidae: Ectatomminae).
The biotechnological potential of venoms to produce drugs with different applications such as antimicrobials, analgesics, antihypertensives, and insecticides has motivated studies of this kind of products . The use of substances obtained from natural products such as animal venoms, including those from insects, is a practice adopted in folk medicine by numerous native populations since ancient times. They would find the raw materials for these therapeutic resources in nature [2,3] for the treatment of the most varied diseases . This is due to the immunological, analgesic, antimicrobial, diuretic, anesthetic, and antirheumatic properties of these products , which also include several substances with potential in the treatment of cancer, microbial, and viral infections, as well as of possible biocide properties such as those of insecticides [5, 6].
Therefore, the search for substances with biological activity obtained from natural products has been an increasingly valuable alternative to find and produce bioactive compounds with practical application [1, 9, 10]. This has motivated numerous recent scientific studies supported by the pharmaceutical industry to search for new bioactive molecules [6,11].
A series of bioactive molecules that are important for several fields such as allergy, immunology, microbiology, biotechnology have already been identified in the venoms of Hymenoptera. Proteins like phospholipases, hyaluronidases, and proteinases have been observed among the identified components, and peptides with diverse activities have already been found in ants, wasps, and bees [1,12].
Thanks to the richness and diversity of bioactive compounds in their venom, Hymenoptera have been gaining notoriety over the last decades, through research studies that use diverse techniques of characterization and biochemical analysis . Many substances of pharmacological interest have been discovered, along with information regarding their structures, biological activities, and action mechanisms. These substances, which vary in diversity and complexity, include pheromones, hormones, defense substances, enzymes, and peptides [1,12,13].
Nevertheless, these insects can cause physiological damage and serious pathologies such as allergies, intense pain, intense sweating, nausea, tachycardia, among other manifestations [14, 15], which in some cases may lead to death of the affected individual . Although the amount of ant venoms produced is small, when compared to that produced by bees and wasps, there is a much larger structural diversity of its components [14, 17]. In light of this, an in depth study of the venoms of Hymenoptera is fundamental as their complex composition and diversity of functions suggest an enormous biotechnological potential for the synthesis of new macromolecules that canbe used for therapeutic purposes or even pest control .
Among the studies on ants,  pointed out the presence of protein allergens with posttranslational modifications in the venom of the Australian ant Myrmecia pilosula. Touchard and collaborators [20,21] have revealed the profiles of venom proteins and peptides of different species of ants through mass spectrometry and high performance liquid chromatography (HPLC), thus reinforcing the thesis that this group of Hymenoptera is promising in the search for molecules to produce new drugs and bioinsecticides. According to Schmidt, Blum, and Overal , E. tuberculatum has one of the most toxic venoms among ants. Pluzhnikov and collaborators and Arseniev [23, 24] have confirmed the high toxicity of its raw venom and have studied the action of the isolated peptide ectatomin. In order to identify the chemical compositionofthe venomof E. tuberculatum in greater detail, it is moreover extremely important to obtain information on the characteristics of its components and their mechanism of action.
In the present study, proteins of the Ectatomma tuberculatum venom have been identified by means of 2D PAGE, followed by tandem nanochromatography with mass spectrometry. A total of 48 proteins have been identified and are discussed for their mechanisms of action. This may help to point out some of these components as a natural prototype of bioactive molecule worthy of synthesizing for pharmaceutical application.
2. Material and Methods
2.1. Protein Extraction from the Ant Glands. Workers of the Ectatomminae ant E. tuberculatum have been collected in the experimental fields of the Cacao Research Center (CEPLAC) at Ilheus, Bahia, Brazil. Additional information on the biology of this species in cacao plantations can be found in Delabie and Hora and collaborators [26, 27]. In the laboratory, 100 workers were anesthetized with C[O.sub.2] and dissected for extraction of venom glands.
The reservoirs of these venom containing glands were removed from the sting apparatus by pulling them out of bodies with forceps and microscissors. The venom reservoirs were then carefully washed and suspended in small volumes of a phosphate buffered saline solution with pH 7.8, punctured, and centrifuged at 10.000x g for 10 min at 4[degrees]C. The supernatants were collected and maintained at -20[degrees]C until use. Venom proteins were extracted from the supernatants according to Rocha and collaborators  and the 2D Quant Kit (GE Healthcare) was used to quantify the proteins. Bovine serum albumin (BSA) was used as standard.
2.2. Two-Dimensional Electrophoresis (2D PAGE). For each sample, 350 [micro]g of venom proteins homogenized in rehydration buffer (7 molx[L.sup.-1] urea, 2 molx[L.sup.-1] thiourea, 1% CHAPS, and traces of bromophenol blue) have been used with DTT 40 mmolx[L.sup.-1] and 0.5% ampholytes at pH 3-10NL with a final volume of 250 [micro]L.
The samples were subjected to IEF in 13 cm gel strips, pH gradient pH 3-10NL (Amersham Biosciences, Immobiline[TM] Dry-Strip). The IEF in the Ettan IPGphor 3 (GE Healthcare) system was carried out according to the manufacturer's manual. The 2D Electrophoresis was performed on 12.5% gel in a SE600 Ruby Vertical Electrophoresis System (Hoefer). The running conditions were similar to those used by Mares and collaborators , staining in 0.08% colloidal Coomassie  for 5 days, followed by destaining in distilled water for 5 days with daily water replacements to remove excess of colloidal Coomassie.
The gels were scanned on ImageScanner II (GE Healthcare) for imaging. Afterwards, it was analyzed using Image Master 2D Platinum 7.0 (GE Healthcare) software to evaluate the number of spots, isoelectric point (pI), molecular mass (MM), and relative abundance of spots on the gels.
2.3. Spot Preparation and MS/MS Analysis. All spots on the gels were excised with the aid of a scalpel, inserted separately into microtubes, and processed according to Silva and collaborators .
After peptide digestion and extraction, 4 [micro]L of the sample corresponding to each spot was applied to the nanoAcquity UPLC (WATERS[R]) and subjected to a C18 "trapping" column of 5 [micro]m, 180 [micro]m x 20 mm, followed by the analytical column of 1.7 [micro]m, 100 [micro]m x 100 mm, under a flow of 0.6 [micro]L/min in a 50 minute run. The peptides were separated according to a gradient of 1% acetonitrile for 1 minute, from 1% to 50% for 40 minutes, from 50% to 85% for 5 minutes, remaining at that concentration for another 2 minutes, then returning to a concentration of 1% in one minute, and remaining in this condition for 2 minutes, totaling 5 minutes of run.
Separated peptides were transferred to the mass spectrometer (Micromass Q-TOF micro), and ionized in a capillary under 3000 V voltage. Next, they were fragmented in positive mode with selection of the minimum intensity of 10 counts, and the 3 of the most intense ions were analyzed by each scan of 1 second, with collision energy ranging from 20 to 95 eV according to the mass/charge (m/z) ratio of the peptides.
The obtained spectra were analyzed by ProteinLynx Global Server 4.2 (WATERS), as well as by MASCOT (Matrix Science), and compared with the SWISSPROT and NCBI databases, respectively. In trypsin hydrolysis, the possible loss of a cleavage site was considered. The tolerance of the peptide masses was [+ or -] 0.3 Da, and the mass tolerance of the fragments was [+ or -] 0.1 Da.
The peptides identified through the NCBI database had their complete sequence located on the same bank and analyzed with NCBI Blast2GO using the BlastP algorithm for protein identification.
Since the E. tuberculatum ant genome has not yet been sequenced, the identification ofthe proteins was based on the complete genome sequences of organisms phylogenetically close to the species.
The 350 [micro]g mass of E. tuberculatum venom protein resolved on 2D PAGE resulted in a well-resolved, low-drag spot profile in the range of 14 to 97 kDa and pH of 4 to 9. A higher density of spots appears for the pH range of 5 to 8.5 and molecular weight of 25 to 93 kDa (Figure 1).
Gels analysis in the ImageMaster 2D Platinum 7.0 software showed the number of spots, the pI and molecular mass values, and the spots' expression levels in each replicate. A total of 244 spots were detected in the gels.
From a total of 129 spots treated with trypsin and analyzed by mass spectrometry, 48 proteins have been identified in the venom of E. tuberculatum. Among these, six showed similarity with not yet characterized proteins.
Table 1 shows the proteins identified with their respective access codes in the NCBI, molecular mass (MM), isoelectric point (pI), score, and peptide sequence for each spot. E. tuberculatum proteins showed sequences similar to the proteins of other Hymenoptera, including wasps, bees, and ants.
According to the Blast2Go software, 42 proteins identified in E. tuberculatum venom can be grouped basically into proteins involved in insect metabolism (annexin B9, nucleoside diphosphate kinase; aconitate hydratase; superoxide dismutase; triose phosphate isomerase; glyceraldehyde-3phosphate dehydrogenase; succinyl-CoA ligase; fructosebisphosphate aldolase; arginine kinase; citrate synthase; enolase; ATP synthase; 4-hydroxybutyrate coenzyme A transferase; medium-chain specific acyl-CoA dehydrogenase, and isocitrate dehydrogenase); transport protein (transferrin); and structural proteins (cyclophilin, actin, and chitinase).
Three proteins found in the venom ofE. tuberculatum are part of a group of compounds with known allergenic effects: triose phosphate isomerase, arginine kinase, and cyclophilinlike protein. Through various techniques of molecular biology, including proteomics studies, allergenic proteins present in Hymenoptera venoms have been identified, as well as their respective action mechanisms.
The triose phosphate isomerase protein, identified in spot 9, relates to the metabolism of carbohydrates, acting as an important enzyme of the glycolytic pathway. It has been studied and classified by  as allergenic because it appeared in the serum of patients presenting allergy to bites of the midge Forcipomyia taiwana (Ceratopogonidae). Also Hoppe, Steinhart, and Paschke  found allergenic enzymes in the protein extract of lychee fruit (Litchi chinensis, Sapindaceae) with high homology with triose phosphate isomerase.
Arginine kinase, expressed in spots 21, 22, 23, 24, and 25 as isoforms of the same protein, is an enzyme that catalyzes the reversible transfer of the phosphate group from ATP to supply the need for energy-requiring cells. It was found in the venom of solitarywasps Eumenidae Eumenes pomiformis and Orancistrocerus drewseni , in the venom of social wasps Polybia paulista (Vespidae), with evidence of its allergenic action in humans  as well as in various other invertebrates such as Dermatophagoides pteronyssinus (Acari: Pyroglyphidae), Blatella germanica (Insecta: Blattodea: Blattellidae), Penaeus monodon (Crustacea: Penaeidae), Homarus gammarus (Crustacea: Nephropidae), Mytilus edulis (Mollusca: Bivalvia: Mytilidae), and Plodia interpunctella (Insecta: Lepidoptera: Pyralidae), a new class of allergens . Yamamoto and collaborators  have isolated arginine kinase from the venom of solitary wasp Pompilidae Cyphononyx dorsalis and confirmed its paralytic activity in spiders, their natural prey. According to Hofling and Rocha , Hymenoptera use their venoms to paralyze or kill their prey, as well as to defend themselves when they feel threatened by vertebrates, including humans. According to , E. tuberculatum is an active predator of cacao pests, feeding on Chrysomelidae the conserved domains to which these proteins belong have been identified, classifying them into specific superfamilies. Spots 6 (SINV.05483), 7 (EAL1007), 8 (L0C.100743840), 64 (L0C_100743840), and 73 (EAL10007) belong to the superfamily LPM010 (Lytic polysaccharide monooxygenase, cellulose degrading). They form a recently discovered group of oxidative enzymes that carries the important role of degrading polysaccharides such as chitin and cellulose, cleaving the glycoside bonds with the oxidation of the C-1, C4 carbons, or both [56, 57]. Spot 77 (SINV.07147) belongs to the superfamily GMC-OXRED-C (glucose-methanolcholine oxidoreductase). They aggregate homologous proteins with oxidation-reduction enzymatic activity, which uses FAD as a cofactor [58, 59].
Some proteins found in the venom ofE. tuberculatum are not part of the protein composition generally found in the venoms of Hymenoptera. It is likely that these proteins are components of the membrane cells of the venom reservoir. Table 2 summarizes the situation for these proteins and their corresponding functions.
The results of the proteomic analysis of the venom of E. tuberculatum allowed the identification of nine proteins related to the process of attack/defense and maintenance of the colony. The active components of the venom have been classified into five subgroups:
(1) Colony asepsis--transferrin and annexin B9: they have antioxidant and antimicrobial activity and are used by the insects to prevent infections of stored food and colony members themselves [34, 53, 54].
(2) Protein of the venom constituents--superoxide dismutase and cyclophilin-like protein: they relate to the preservation of the functional integrity of cellular/glandular constituents, preventing oxidation and damage to the cellular structure of the venom bag and its reservoir [35, 40, 41,44].
(3) Diffusion of the venom in the prey--chitinase, enolase, annexin B9, and actin: the process of diffusion of the constituents of the venom .
(4) Prey paralysis--arginine kinase: it causes neurotoxicity, leading to paralysis and subsequent death of prey .
(5) Alteration of homeostasis and cellular toxicity--actin, enolase, glyceraldehyde-3-phosphate dehydrogenase: they cause damage to different tissue types and alteration of cellular homeostasis and maybe neurotoxic .
Figure 2 resumes the scheme for the aforementioned functions.
The identification and characterization of the proteins present in the venom of E. tuberculatum can aid in the better understanding of how the ants use the protein constituents for different behaviors such as defense against natural and pathogenic enemies, attack on prey, and maintenance of the colony, in addition to correlating the presence of certain allergic components in the venom with effects occurring in humans after inoculation. Furthermore, for providing knowledge as explained above, studies like this one also contribute to the identification of substances as possible prototypes for synthesis of macromolecules for new purposes, from their use as molecular markers to the manufacture of drugs and insecticides.
Conflicts of Interest
The authors declare that they have no conflicts of interest.
Special thanks are due to Martin Brendel for reviewing the English manuscript. This research received support from the PRONEX Program [SECTI-FAPESB & CNPq, project 011/2009]. Finally JRS acknowledges her study (MS) grant from CAPES and JHCD acknowledges his research grant from CNPq.
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Juliana Rocha da Silva, (1) Aline Zanotelli de Souza, (2) Carlos Priminho Pirovani, (2) Helena Costa, (2) Aline Silva, (2) Joao Carlos Teixeira Dias, (2) Jacques Hubert Charles Delabie, [ID](2,3) and Renato Fontana [ID] (2)
(1) Universidade Estadual de Santa Cruz (UESC), Programa de Pos-Graduacao em Biologia e Biotecnologia de Microrganismos (PPGBBM), 45.662-900 Ilhous, BA, Brazil
(2) Universidade Estadual de Santa Cruz (UESC), Ilheus, BA, Brazil
(3) Laboratorio de Mirmecologia, Centro de Pesquisas do Cacau (CEPLAC), Itabuna, BA, Brazil
Correspondence should be addressed to Renato Fontana; email@example.com
Received 22 November 2017; Revised 20 March 2018; Accepted 29 May 2018; Published 21 June 2018
Academic Editor: Leonardo Dapporto
Caption: Figure 1: Spots corresponding to the proteins identified in the bidimensional gel of Ectatomma tuberculatum venom. Isoelectric point (pI) of 3 to 10, molecular weight (MW) of 14.4 to 97.0 kDa. The numbered spots correspond to proteins identified by mass spectrometry.
Caption: Figure 2: Representation of the action mechanisms proposed for the proteins identified in the venom of E. tuberculatum; 'peptide ectatomin [23, 25] not identified in the present study.
Table 1: Proteins identified in the Ectatomma tuberculatum venom with their respective access codes in NCBI, molecular mass (MM), isoelectric point (pI), score, and peptide sequence of each spot were analyzed in the Mascot program (Matrix Science). Spot Protein/Source Accession code NCBI 01 Annexin B9/Harpegnathos gi|307194598 saltator (Formicidae) 76 Annexin B9/Harpegnathos gi|307194598 saltator (Formicidae) 02 Nucleoside diphosphate gi|307173082 kinase/Camponotus floridanus (Formicidae) Aconitate hydratase, mitochondrial/ 03 Acromyrmex echinatior gi|332018721 (Formicidae) Aconitate hydratase, mitochondrial/ 51 Acromyrmex echinatior gi|332018721 (Formicidae) 52 Aconitate hydratase, gi|332018721 mitochondrial/ Acromyrmex echinatior (Formicidae) 04 Superoxide dismutase gi|307204104 [Cu-Zn]/Harpegnathos saltator (Formicidae) 05 Cyclophilin-like protein- gi|292397870 Nylanderia nr. pubens LZ- 2010 (Formicidae) 09 Triosephosphate gi|332024520 isomerase/Acromyrmex echinatior (Formicidae) 12 Phosphoglycerate mutase gi|340726229 2-like/Bombus terrestris (Apidae) 16 Glyceraldehyde-3- gi|307181618 phosphate dehydrogenase/ Camponotus floridanus (Formicidae) 17 Glyceraldehyde-3- gi|307181618 phosphate dehydrogenase/ Camponotus floridanus (Formicidae) 18 Glyceraldehyde-3- gi|307181618 phosphate dehydrogenase- Camponotus floridanus (Formicidae) 69 Glyceraldehyde-3- gi|332026368 phosphate dehydrogenase 2/ Acromyrmex echinatior (Formicidae) 19 Succinyl/CoA ligase [GDP/ gi|307170861 forming] subunit alpha, mitochondrial/Camponotus floridanus (Formicidae) 26 Succinyl/CoA ligase [ADP/ gi|307178345 forming] subunit beta, mitochondrial / Camponotus floridanus (Formicidae) 57 Succinyl/CoA ligase [GDP/ NCBI gi|307170861 forming] subunit alpha, mitochondrial/Camponotus floridanus (Formicidae) 20 Fructose/bisphosphate gi|110748949 aldolase-like/Apis mellifera (Apidae) 58 Fructose/bisphosphate gi|110748949 aldolase/like /Apis mellifera (Apidae) 75 Fructose-bisphosphate gi|307206615 aldolase/ Harpegnathos saltator (Formicidae) 78 Fructose-bisphosphate gi|307206615 aldolase/ Harpegnathos saltator (Formicidae) 21 Arginine kinase / gi|332018357 Acromyrmex echinatior (Formicidae) 22 Arginine kinase / gi|332018357 Acromyrmex echinatior (Formicidae) 23 Arginine kinase / gi|307197996 Harpegnathos saltator (Formicidae) 24 Arginine kinase / gi|332018357 Acromyrmex echinatior (Formicidae) Arginine kinase / 25 Acromyrmex echinatior gi|332018357 (Formicidae) 27 Actin-5, muscle-specific/ gi|307197034 Harpegnathos saltator (Formicidae) 29 Probable citrate synthase gi|307202019 1, mitochondrial/ Harpegnathos saltator (Formicidae) 30 Probable citrate synthase gi|350425352 2, mitochondrial/ Bombus impatiens (Apidae) 34 ATP synthase subunit beta gi|307195440 mitochondrial/ Harpegnathos saltator (Formicidae) 32 ATP synthase subunit gi|269784695 alpha, mitochondrial precursor/ Nasonia vitripennis (Pteromalidae) 31 Enolase/ Harpegnathos gi|307211488 saltator (Formicidae) 33 Enolase/ Harpegnathos gi|307211488 saltator (Formicidae) 61 Enolase/ Harpegnathos gi|307211488 saltator (Formicidae) 35 4-hydroxybutyrate gi|307174077 coenzyme A transferase/ Camponotus floridanus (Formicidae) 37 Chitinase-like protein gi|307207611 Idgf4-Harpegnathos saltator (Formicidae) 60 Probable medium-chain gi|156553409 specific acyl-CoA dehydrogenase, mitochondrial-like/ Nasonia vitripennis (Pteromalidae) 66 Transferrin/ Harpegnathos gi|307215135 saltator (Formicidae) 71 Transferrin/ Acromyrmex gi|332029256 echinatior (Formicidae) 72 Transferrin/ Acromyrmex gi|332029256 echinatior (Formicidae) 74 Transferrin/Acromyrmex gi|332029256 echinatior (Formicidae) Probable isocitrate dehydrogenase [NAD] subunit alpha, mitochondrial/ 70 Camponotus gi|307166568 floridanus (Formicidae) 06 Uncharacterized protein gi|322778685 SINV.05483/ Solenopsis invicta (Formicidae) Uncharacterized protein 07 EAL10007/ Harpegnathos gi|307206139 saltator (Formicidae) Uncharacterized protein 73 EAL10007/ Harpegnathos gi|307206139 saltator (Formicidae) 08 Uncharacterized protein gi|350427541 LOC100743840/ Bombus impatiens (Apidae) 64 Uncharacterized protein gi|350427541 L0C100743840/Bombus impatiens (Apidae) Uncharacterized protein 77 SINV_07147/Solenopsis gi|322796407 invicta (Formicidae) Spot MM/pI Score 01 35836/4.93 68 76 35836/4.93 106 02 19547/8.41 68 03 92836/8.65 101 51 92836/8.65 371 52 92836/8.65 406 04 14008/6.18 218 05 18008/8.89 162 09 26935/7.71 129 12 35071/7.17 90 16 35815/7.71 133 17 35815/7.71 194 18 35815/7.71 174 69 37576/8.15 81 19 35682/8.93 108 26 48972/6.63 125 57 35682/8.93 158 20 39975/7.57 85 58 39975/7.57 85 75 40185/8.31 264 78 40185/8.31 338 21 40032/5.86 370 22 40032/5.86 238 23 39996/5.75 447 24 40032/5.86 333 25 40032/5.86 333 27 42098/5.30 114 29 49316/8.94 85 30 51596/9.03 72 34 55223/5.32 397 32 59247/9.18 131 31 47379/5.79 276 33 47379/5.79 230 61 47379/5.79 400 35 53245/7.99 104 37 55766/8.45 56 60 45544/8.37 67 66 82014/5.66 87 71 79357/5.47 145 72 79357/5.47 431 74 79357/5.47 84 70 29723/6.01 74 06 17131/6.70 123 07 27511/6.70 74 73 27511/6.70 85 08 24614/6.41 72 64 24614/6.41 78 77 68188/6.51 97 Spot Peptide sequences 01 GIGTTDSTLIR 76 GIGTTDSTLIR 02 TFIMVKPDGVQR, VMLGETNPKDSAPGTIR 03 SEIAGAADQHK, SAFNVTPGSEQIR, NQLTNEWGAVPDVAR 51 LDFNPVKDR, NQLTNEWGAVPDVAR, VAVPSTIHCDHLIEAQIGGNEDLQR 52 LDFNPVKDR, SEIAGAADQH, NQLTNEWGAVPDVAR 04 LACGVIGITK, TLVVHADPDDLGQGGHELSK 05 VFFDMTADDKPVGR, VIPNFMCQGGDFTNHNGTGGK 09 IIYGGSVTAANAK, VIACIGEKLEER 12 FLGDEETVK, IIIAAHGNSLR, YGEEQVQIWR, YADGPKPEEFPK 16 VKEASEGPLK, VPVHNVSVVDLTVR, VVSNASCTTNCLAPLAK 17 VKEASEGPLK, IAVFSEREPK, LAKPASYDAIK, VPVHNVSVVDLTVR 18 VKEASEGPLK, IAVFSEREPK, LAKPASYDAIK, VPVHNVSVVDLTVR 69 IAVFSEREPK 19 LLEQNKSR, SPAQMGNELLK, GGAQDKINALEK, KAGTEHLGKPVFK, QGTFHCQQAIDYGTK, LIGPNCPGIIAPEQCK 26 FDDNAEFR, MCETPEEAK, ICNAVMVTQR, ICNAVMVTQR, IVPIDDLDEAAR, LHGGEPANFLDVGGGASASAVK 57 LIGPNCPGIIAPEQCK 20 LAILENANVLAR 58 LAILENANVLAR 75 GILAADESTATIGK, GILAADESTATIGKR 78 GILAADESTATIGK, GILAADESTATIGKR 21 FLQAANACR, LVTAVNEIEK, LGLTEYQAVK, GTFYPLTGMSK, EMNDGIAELIK, VSSTLSGLTGELK, EGDRFLQAANACR, LIDDHFLFKEGDR, IISMQMGGDLGQVYR, GTRGEHTEAEGGIYDISNK, GTRGEHTEAEGGIYDISNK LIDDHFLFK, EMNDGIAELIK, 22 LVTAVNEIEKR, VSSTLSGLTGELK, IISMQMGGDLGQVYR, GTRGEHTEAEGGIYDISNK, LGLTEYQAVKEMNDGIAELIK, GIFHNDDKTFLVWCNEEDHLR 23 LGLTEYQAVK, LIDDHFLFK, IISMQMGGDLGQVYR, SLDGYPFNPCLTEAQYK, GIFHNDDKTFLVWCNEEDHLR LVTAVNEIEK, LGLTEYQAVK, 24 LIDDHFLFK, EMNDGIAELIK, LVTAVNEIEKR, IISMQMGGDLGQVYR, IISMQMGGDLGQVYR LVTAVNEIEK, LGLTEYQAVK, LVTAVNEIEKR, LIDDHFLFKEGDR, 25 IISMQMGGDLGQVYR, GTRGEHTEAEGGIYDISNK, GTRGEHTEAEGGIYDISNK 27 GYSFTTTAER, HQGVMVGMGQK, DSYVGDEAQSKR, QEYDESGPGIVHR, VAPEEHPVLLTEAPLNPK, DLYANNVLSGGTTMYPGIADR 29 AISQEWAS, SGQVVPGYGHAVLR, VGEVTVDMMYGGMR 30 VVPPILLETGK, VGEVTVDMMYGGMR 34 VVDLLAPYAK, TIAMDGTEGLVR, AHGGYSVFAGVGER, FTQAGSEVSALLGR, VALVYGQMNEPPGAR, LVLEVAQHLGENTVR, IMDPNIIGMEHYNIAR, IPSAVGYQPTLATDMGTMQER 32 HALIIYDDLSK, TAiAIDTIINQKR, EAYPGDVFYLHSR, VVDALGNPIDGKGPLNNK 31 MGSEVYHYLK, SNGWGTMVSHR, IGMDVAASEFY 33 MGSEVYHYLK, IGMDVAASEFYK, VNQIGSVTESINAHK 61 IGMDVAASEFYK, VNQIGSVTESINAHK, AAVPSGASTGVHEALELR 35 IVGSFCVGSEK, IQPVLTSGAGVVTNR 37 EADYPAPIYGSYGR 60 TNPDPKAPASK 66 DLDINNVQGLR 71 YEAVAVIHK, DNGADITIIDGGSVK, FDCILEKDEAACLK, LTAMGVLTDINNPEYSAR 72 DNGADITIIDGGSVK, LTAMGVLTDINNPEYSAR 74 DNGADITIIDGGSVK 70 VAEFAFKYATDNNR 06 YPLPLADGSGYK, ELETDECFNKYPLPLADGSGYK 07 GAMGCGPQETFR 73 GAMGCGPQETFR 08 YGTGVIVQR 64 YGTGVIVQR 77 VYKDLPVGK Table 2: Proteins found in the venom of E. tuberculatum with unknown function in the venoms of Hymenoptera. Protein Spot Medium-chain specific acyl-CoA 60 dehydrogenase, mitochondrial-like (E. C. 22.214.171.124) Aconitate hydratase mitochondrial 3, 51 and 52 (E.C. 126.96.36.199) (isoforms) ATP Synthase (E.C. 188.8.131.52) 32 Citrate Synthase (E.C. 184.108.40.206) 29, 30 and 34 Fructose-bisphosphate aldolase 20, 58, 75 and 78 (E.C. 220.127.116.11) Glyceraldehyde-3-phosphate 16,17,18 and 69 dehydrogenase (E.C. 18.104.22.168), Isocitrate dehydrogenase (E.C. 70 22.214.171.124) Nucleoside diphosphate kinase 2 (E.C. 126.96.36.199) Phosphoglycerate mutase 2-like 12 (E.C. 188.8.131.52) Succinyl-CoA ligase (E.C. 184.108.40.206) 19, 26 and 57 4-Hydroxybutyrate coenzyme A 35 transferase (E.C. 220.127.116.11) Protein Action Mechanism Medium-chain specific acyl-CoA Enzyme that acts on the [beta]- dehydrogenase, mitochondrial-like oxidation of fatty acids in (E. C. 18.104.22.168) mitochondria. Aconitate hydratase mitochondrial Catalyzes the isomerization of (E.C. 22.214.171.124) citrate in isocitrate in the citric acid cycle. Some studies have revealed the presence of this protein in the venoms of several arthropods, such as Vespidae Vespula maculifrons, Sicariidae Loxosceles reclusa spider, A. mellifera, among others, as well as in the cuticle of Hemiptera Coccidae Ericeruspela [7, 8]. ATP Synthase (E.C. 126.96.36.199) ATP-forming enzyme complex from ADP and inorganic phosphate during oxidative phosphorylation in respiration and photosynthesis processes. Citrate Synthase (E.C. 188.8.131.52) Participates in the first step of the citric acid cycle by catalyzing the condensation reaction of an acetate residue containing two carbons of an acetyl coenzyme A with an oxaloacetate molecule containing four carbons to form a six- carbon citrate. Fructose-bisphosphate aldolase Involved in carbohydrate (E.C. 184.108.40.206) metabolism, and responsible for the cleavage of fructose 1,6- bisphosphate in two trioses: glyceraldehyde 3-phosphate and dihydroxyacetone phosphate in glycolysis. Glyceraldehyde-3-phosphate Participates in glucose dehydrogenase (E.C. 220.127.116.11), metabolism by the Embden- Meyerhof route, catalyzing the conversion of glyceraldehyde 3- phosphate to 1,3- diphosphoglycerate. Isocitrate dehydrogenase (E.C. Enzyme that catalyzes oxidative 18.104.22.168) decarboxylation of isocitrate to form a-ketoglutarate in the citric acid cycle. Nucleoside diphosphate kinase Participates in nucleotide (E.C. 22.214.171.124) biosynthesis, catalyzing the conversion of the nucleoside diphosphate into nucleoside trisphosphate. Phosphoglycerate mutase 2-like Glycolytic enzyme which (E.C. 126.96.36.199) catalyzes the conversion of 3- phosphoglycerate to 2- phosphoglycerate Succinyl-CoA ligase (E.C. 188.8.131.52) Enzyme that participates in the citric acid cycle, catalyzing the conversion of succinyl-CoA to succinate. 4-Hydroxybutyrate coenzyme A Enzyme that participates in the transferase (E.C. 184.108.40.206) metabolic process involving acetyl Co-A, exhibiting transferase activity. It is well studied in bacteria and participates in the fermentation process.
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|Title Annotation:||Research Article|
|Author:||da Silva, Juliana Rocha; de Souza, Aline Zanotelli; Pirovani, Carlos Priminho; Costa, Helena; Silva,|
|Publication:||Psyche (Cambridge, 1874)|
|Date:||Jan 1, 2018|
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