SciELO - Scientific Electronic Library Online

 
vol.9 issue2An idea comes true...Interaction between infection, nutrition and immunity in tropical medicine author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand

Journal

Article

Indicators

Related links

Share


Journal of Venomous Animals and Toxins including Tropical Diseases

On-line version ISSN 1678-9199

J. Venom. Anim. Toxins incl. Trop. Dis vol.9 no.2 Botucatu  2003

http://dx.doi.org/10.1590/S1678-91992003000200002 

REVIEW ARTICLE

 

Hymenoptera venom review focusing on Apis mellifera

 

 

P. R. de Lima; M. R. Brochetto-Braga

Department of Biology, Institute of Biosciences of Rio Claro, State of São Paulo, Brazil

Correspondence

 

 


ABSTRACT

Hymenoptera venoms are complex mixtures containing simple organic molecules, proteins, peptides, and other bioactive elements. Several of these components have been isolated and characterized, and their primary structures determined by biochemical techniques. These compounds are responsible for many toxic or allergic reactions in different organisms, such as local pain, inflammation, itching, irritation, and moderate or severe allergic reactions. The most extensively characterized Hymenoptera venoms are bee venoms, mainly from the Apis genus and also from social wasps and ant species. However, there is little information about other Hymenoptera groups. The Apis venom presents high molecular weight molecules - enzymes with a molecular weight higher than 10.0 kDa - and peptides. The best studied enzymes are phospholipase A2, responsible for cleaving the membrane phospholipids, hyaluronidase, which degrades the matrix component hyaluronic acid into non-viscous segments and acid phosphatase acting on organic phosphates. The main peptide compounds of bee venom are lytic peptide melittin, apamin (neurotoxic), and mastocyte degranulating peptide (MCD).

Key words: Hymenoptera, venom, enzymes, peptides.


 

 

INTRODUCTION

HYMENOPTERA VENOM OVERVIEW

Hymenoptera have venom glands producing several chemicals, which are stored in their reservoirs. The venom can be injected through a true sting or a barbed tip that is directed into the victim’s body. All Hymenoptera venoms have protein or peptide elements, many with pharmacological properties (8).

The study of social Hymenoptera (bees, wasps, and ants) venom proteins is of great interest, since these venoms can trigger serious allergenic reactions in humans. Most of these proteins are enzymes, specific toxins, or other bioactive molecules; several of them have been characterized and their primary structures determined by biochemical techniques (28).

There can be toxic (local) or allergenic reactions to Hymenoptera venoms. The former are caused mostly by low molecular weight compounds, which can result in pain, local inflammation (26,28), itching, and irritation as immediate responses that after some hours are attenuated (20). Melittin is the main compound responsible for most of these reactions, and it is present in several bee venoms. However, most of the serious reactions reported in victims stung by Hymenoptera are allergenic, which, in their majority, result from a combination of mastocytes with IgE, triggering a cascade of mediators, including histamine, leucotrienes, platelet activating factors, enzymes, peptides, and other chemicals (26,28). In fact, 50-80% of victims present IgE specific molecules to venom compounds. These reactions cause lasting local inflammation and are called "late phase reactions" (20).

A few people develop serious sensibility to Hymenoptera venoms, and some studies have reported that there is a 25% sensitivity to bee venom. In severe cases, cardiorrespiratory failure or systemic anaphylactic shock (26,49), followed by cutaneous symptoms can occur (20).

Because the risk of sensitization tends to increase with age (20,28), there are many reported fatal cases mainly in adults, although they are uncommon.

The most extensively characterized Hymenoptera venoms are bee venoms, mainly from the Apis genus, and also from social wasps (Vespula, Vespa, and Polistes ) and some ant species (8,42). However, little information is known about venoms from other groups.

Social wasp venoms (Vespidae family) generally cause lasting pain, local edema, and redness due to an increased permeability of the skin blood capillaries. The allergenic reactions in wasp victims can be as serious as those from bees, although less common.

Social wasp venoms, specially in some Vespula, Vespa, and Polistes genera and in Apis bee venom contain a blend of biogenic amine compounds, such as histamine, serotonin, dopamine, noradrenaline, and polyamines as well as several toxins, peptides, and proteins (17,42). The main protein allergens present phospholipase, hyaluronidase, and acid phosphatase activities. Mueller et al. (40) investigated differences in venom protein composition of Vespula maculifrons, Vespula maculate, and Vespula arenaria in relation to enzymatic activities of phospholipase A1B and hyaluronidase and some immunological properties of some allergenic compounds. Hoffman and Jacobson (29) made a comparative study of protein composition from twelve different bee and wasp venoms (genera Apis, Bombus, Xilocopa, Vespula, Dolichovespula, and Polistes) and found quantitative differences.

Knowledge on the composition and properties of ant venoms is still limited, as there is great difficulty in obtaining sufficient venom samples for chemical analysis, and composition varies greatly between the different ant subfamilies. This makes any conclusion too difficult (17,18). The most studied venoms have been from Myrmicinae, Ponerinae, and Formicinae subfamilies; they possess histamine, hyaluronidase, and kinine-like activities (48). Ant venoms are constituted by a simple organic acid mixture as in Formicinae, or even of complex mixtures of proteins and neurotoxins as in Ponerinae and Myrmicinae, like in bees and wasps The enzymes found in ant venoms are the same as currently found in other Hymenoptera families, and are of great importance in allergic processes (17,18).

Apis mellifera VENOM

In the Apidae family (social, solitary bees and bumblebees), little is known about bumblebee venoms, which is in contrast to the extensive information on the Apis mellifera venom, probably the best characterized insect venom (17).

In this family, venom is of high importance in the colony defense and is its primary function (3,15). The main bee enemies are usually also insects, including Hymenoptera, such as ants, wasps, bumblebees, and even bees from other colonies (3). The stinging behavior is most commonly observed near the colony (15), and is mainly triggered by a pheromone secretion (26).

Venom glands in the worker bees become active just after adult emergence and their maximal production is achieved within two or three weeks after the emergence. Venom composition also undergoes some changes during bee lifetime, and these changes are believed to occur mainly due to a changing behavior from the maintenance to food gathering through life. Venom production is also higher during summer months, in which there is a peak of activity in the colony, and when the relatively young individuals are beginning their defense behavior (26).

To the small victims, such as insects, one sting can be fatal, or can provoke intense myotoxic reaction (41); to humans, the sting is merely an unpleasant experience, despite serious cases of allergic reaction. On the other hand, there is a potential therapeutic value in bee venom, particularly in arthritic and rheumatic conditions (3,17). Thus, studies on venom composition and properties, their individual compounds can be useful for a better understanding of these effects, such as eventual therapeutic applications (3).

Some venom compounds have been extensively used in basic research on properties of natural and synthetic membranes (melittin), of smooth muscle nervation (apamin), and secretory and anti-inflammatory mechanisms (mastocyte degranulating peptide) (3).

Apis mellifera venom is composed of high molecular weight molecules - enzymes with a molecular weight higher than 10.0 kDa and peptides (with low molecular weight).

A) Peptides

So far, melittin is the best characterized peptide. It has alkaline characteristics similar to other venom compounds and seems to be the major responsible for intense local pain (17,22). It corresponds to 40-50% of venom dry weight, although this varies during aging (46). Chemically, it is considered one of the most interesting peptides studied to date. It has 26 amino acid residues with amphypathic characteristics (polar and non-polar ends), which allows it to interact with lipid membranes, which in turn, can increase permeability of the erythrocytes and other cell membranes (3,15,22,28,51). Its tetrameric form is believed to have ionophore properties being responsible for a constant skin nerve terminal depolarization that causes pain from the sting (17). Although most experimental works had demonstrated melittin lytic action on erythrocytes, this peptide can also trigger lysis in many cell types and intercellular membranes, such as lysosomes (15,22), with subsequent enzyme release, as demonstrated by Hegner (25). When this author studied isolated leukocyte granules by electron microscopy, he found that they had reduced diameter, membrane fractures, and increased cellular matrix density. Other researchers also found that, depending on its concentration or the organ studied, this peptide can dilate or contract blood vessels, and depolarize and contract heart or skeletal muscles. It can also increase vascular permeability at the sting site and elicit slow contraction of smooth muscles, although its action is transient (15,22). Melittin also presents fungicide and antibiotic action against several microorganisms (22).

Its biosynthesis was studied in vivo by feeding worker and queen bees with radioactive amino acids that were incorporated into the precursor of melittin molecule - the promelittin. This precursor differs from melittin because it has an oligopeptide chain at the amino-terminal position, and differently from the processed or mature molecule, it is not detected in inoculated venom in the victim (34).

Another important peptide in bee venom is apamin, a well-characterized peptide (15,51). Apamin is a small peptide that corresponds to less than 2% of venom dry weight. It has about 2.0 kDa, only 18 amino acid residues, and neurotoxic properties. It was firstly reported as an inductor of convulsions in mice (24,43), but this compound does not exert any influence on a great variety of mammal cells (26). Initially, it was believed that this peptide had specific effect on synaptic functions in the central nervous system. However, it is now known that apamin affects Na+ and Ca++ channels in several cells and does not present lytic properties (15,17,51). Similar to many potent venom neurotoxins, apamin binds with high affinity to specific receptors of a post-synaptic membrane and seems to block many inhibitory or hyper-polarization effects, including a-adrenergic, colinergic, purinergic, and produce the relaxing of the neurotensin-induced effects. These actions are due to the blocking of the post-synaptic ion channels, which have an important role in repetitive activities in neurons of both vertebrates and invertebrates; consequently, apamin blocks these activities (26).

In contrast to melittin, apamin action mechanism cannot be explained only by its chemical structure. In 1981, through crystallographic methods, Hider and Ragnarson (27) determined the secondary structure of apamin and made different structural comparisons and correlations among the peptides of the venom.

A few years ago, a recombinant form of apamin was produced by molecular biology techniques (14). In the same year, Deschaux and coworkers (13) found that apamin plays a role in the rat learning process, acting directly on the central nervous system.

The MCD peptide (mastocyte degranulating peptide), chemically similar to apamin, is probably the main venom factor responsible for a massive release of histamine (17). It represents about 2% of the venom dry weight (19). Most experiments with MCD were performed with rat peritoneal cells (3). This peptide structure was determined by Dotimas and collaborators (16) by chromatographic techniques and spectral studies. The MCD has 22 amino acid residues of about 3.0 kDa (19), is rich in a-helix, and presents two disulfide bonds in its structure (26). Some years ago, the MCD and apamin genes were found to share a same exon (19). As the own name suggests, the MCD presents a degranulating mastocyte property; at low concentrations, it causes a release of a large amount of histamine (15,51). Like apamin, this peptide probably binds to specific protein membrane receptors, like apamin (26). The mastoparans present in the venoms of some wasp species also have some degranulating mastocyte properties at high concentrations.

Histamine molecules, able to induce pain in mammals, are also present in bee venom, although their concentration is very low (26).

Apart from these main compounds, there is a great diversity of low molecular weight (up to 10.0 kDa) chemical compounds at low concentrations, which have not been well characterized (3,15,17,48). For instance, was found in bee venom, a serine-protease inhibitor with a molecular weight of 9.0 kDa (54).

b) Enzymes

Enzymes represent the high molecular weight fraction of the venom (15.0 to 50.0 kDa). The major Hymenoptera venom enzyme is the phospholipase A2 (PLA2), which is also the most studied in bee venom (51). In snake venoms, this enzyme exists as several isoenzymes, with many physiological actions, such as cardiotoxic, neurotoxic, myotoxic, anticoagulant activity, and edema induction (44). The PLA2 from bee venom has a molecular weight of about 15.0-16.0 kDa. However, this enzyme can also present a molecular weight of 19.5 kDa when a carbohydrate residue is bound, or even 38.5 kDa when in analytical centrifugation. It is described as a potent bee venom allergen. It represents about 12% of the crude venom and it is extremely alkaline. It has the interesting cleavage property of the main construction block of biological membranes – the phospholipids (phosphatidylcholine, for instance), producing lisophospholipids and long chain anionic fatty acids. It causes pores in the membrane, and consequently, cellular lysis (3,15,28,51). It can be responsible for a series of indirect venom pharmacological reactions (51). This enzyme has been extensively studied in bee venom, and its action and kinetic properties have been determined (1,2). Many studies have already shown a synergistic reaction of phospholipase A2 with melittin in mammal erythrocyte lysis process (6,57), and the authors reported that melittin facilitates the exposure of membrane phospholipids to the catalytic site of the enzyme, by opening melittin-induced channels. In immunological terms, a short time ago, the carbohydrate residues present in the bee venom PLA2 molecule was, a short time ago, believed to play an essential role in the specific IgE induction in sting victims. However, Okano et al. (45) demonstrated that these residues have low significance in relation to the deglycosilated molecule in in vivo IgE synthesis. A toxin with phospholipase A2 activity was isolated from the social wasp Agelaia pallipes pallipes venom by Costa and Palma (12). This enzyme presented a very potent hemolytic activity in monomeric form.

Hyaluronidase (HYAL) is the enzyme responsible for hyaluronic acid hydrolysis and condroitin sulfate, both abundant in connective tissue, mainly in the interstitial space. HYAL is known as the "spreading factor", since it degrades the hyaluronic acid to non-viscous segments, allowing the fast spreading of the venom compounds through the interstitial space (15). The same way as PLA2, HYAL has varied molecular weight from 30.0 to 60.0 kDa, depending on the carbohydrate quantity bound to the protein, and it presents an optimal activity between pH 4.0 and 5.0 (3,15,51). Kemeny et al. (32) purified and characterized the enzyme by chromatographic methods as an alkaline glycoprotein rich in aspartic and glutamic acid (with its amino-terminal end blocked). Its activity has been detected in bee and snake venoms and has been purified by electrophoretic and chromatographic methods (35,47). Bee venom enzyme presents asparagine-linked carbohydrates that are mostly oligossaccharides (35).

The acid phosphatase enzyme (AcP) or phosphomonoesterase was firstly described by Benton (5). This enzyme represents approximately 1% of the venom dry weight (3). Significant amounts of this enzyme have already been found in bee venom as dimmers of a protein chain with about 49.0 kDa. Its optimal pH is between 4.4-4.8. It is allergenic, although its biological function is still unclear in Hymenoptera venoms (3,4,28). Purified samples of this enzyme also revealed a glycoprotein nature, the same way as PLA2 and HYAL (4). AcP is a potent releaser of histamine in human basophils, thus relevant in allergic process to the venom (58). Therefore, the study on the properties of this enzyme has significant importance to the understanding of the venom allergic properties (4).

In bee venom, there are still reports of the existence of a-glycosidase, lysophospholipase, esterase, and lipase activities; the two latter being involved in the cell lysis process, although their specific functions are still unclear (51). The a-glycosidase enzyme represents 0.6% of the crude venom dry weight with an optimal pH of about 5.5. It seems that this enzyme is associated with honey production by bees. The lysophospholipase is likely to act increasing PLA2 activity. While PLA2 degrades the phospholipids into lisophospholipids, these latter are cleaved by the lysophospholipases into glycerophosphocoline and anionic fatty acid (3).

In insect venom there is very little information about their proteases (10,11,36,37,51,52) since low levels of activity were detected. However, they have already been detected in bee and other social Hymenoptera venoms, being responsible for moderate necrosis in some tissues (52).

As an exception to that observed in most studies on Hymenoptera venoms, high levels of proteolytic activity have already been detected in Bombus pensylvanicus bumblebee venom (30) associated with PLA2, HYAL, and AcP activities. Using sequencing techniques, the authors determined that Bombus pensylvanicus protease has 243 amino acid residues and a molecular weight of 27.0 kDa. The proteolytic activity has also been detected in the venoms of a social wasp species (Polistes infuscatus), an ant species (Eciton burchelli) (53), and Vespa orientalis (23); this latter with anticoagulant properties and molecular weight of about 40.0 kDa. Furthermore, proteolytic activity has also been determined in Polybia paulista, Polybia ignobilis, Agelaia pallipes pallipes, and Apoica pallens venoms, in which several isoenzymes were observed with caseinolytic and gelatinolytic activities (50).

From these, the most extensively studied was the Bombus venom protease, which presented a tryptic amidase specificity with strong allergenic reaction. Other venoms from Bombus species were also analyzed regarding this feature (Bombus impatiens, Bombus fraternus, and Bombus bimaculatus) and also exhibited tryptic amidase activity (30).

Differently from that observed in insect venoms, there is much more information about proteolytic enzymes in snake venoms (9,21,31,33,38,55,56), demonstrating that these enzymes can play different roles, exhibiting coagulant, fibrinolytic (33,39), and hemorrhagic effects (7,39).

 

REFERENCES

1 AHMED T., KELLY SM., LAWRENCE AJ., MEZNA M., PRICE NC. Conformational changes associated with activation of bee venom, phospholipase A2. J. Biochem., 1996, 120, 1224-31.        [ Links ]

2 ANNAND RR., KONTOYIANNI M., PENZOTTI JE., DUDLER T., LYBRAND TP., GELB MH. Active site of bee venom phospholipase A2: the role of histidine-34, aspartate-64 and tyrosine-87. Biochemistry, 1996, 35, 4591-601.        [ Links ]

3 BANKS BEC., SHIPOLINI RA. Chemistry and pharmacology of honey bee venom. In: PIEK T. Venoms of the hymenoptera: biochemical, pharmacological and behavioral aspects. London: Academy Press, 1986: 329-416.        [ Links ]

4 BARBONI D., KEMENY DM., CAMPOS S., VERNON CA. The purification of acid phosphatase from honey bee venom (Apis mellifera). Toxicon, 1987, 25, 1097-103.        [ Links ]

5 BENTON AW. Esterases and phosphatases of honey bee venom. J. Apic. Res., 1967, 6, 91-4.        [ Links ]

6 BERNHEIMER AW., RUDY B. Interactions between membranes and cytolytic peptides. Biochim. Biophys. Acta, 1986, 864, 123-41.        [ Links ]

7 BJARNASSON JB., FOX JW. Hemorrhagic metalloproteinases from snake venoms. Pharmacol. Ther., 1994, 62, 325-72.         [ Links ]

8 BLUM MS. Biochemical defenses of insects. In: ROCKSTEIN M. Biochemistry of insects. London: Academic Press, 1978: 465-513.        [ Links ]

9 BOLAÑOS R. Toxicity of costa rican snake venoms for the white mouse. Am. J. Trop. Med. Hyg., 1972, 21, 360-3.        [ Links ]

10 BROCHETTO-BRAGA MR., CHAUD-NETO J., LIMA PRM., RODRIGUES A., CARBONE SC. Biochemical characterization of venom proteases from different honeybee races. In: REUNIÃO ANUAL DA SOCIEDADE BRASILEIRA DE BIOQUÍMICA E BIOLOGIA MOLECULAR (SBBq), 24, Caxambú, 1995. Anais... Caxambú: Sociedade Brasileira de Bioquímica e Biologia Molecular, 1995. 128.        [ Links ]

11 BROCHETTO-BRAGA MR., CHAUD-NETO J., LIMA PRM., RODRIGUES A. Enzymatic variability of venoms from Apis mellifera subspecies. In: REUNIÃO ANUAL DA SOCIEDADE BRASILEIRA DE BIOQUÍMICA E BIOLOGIA MOLECULAR (SBBq), 25, Caxambú, 1996. Anais... Caxambú: Sociedade Brasileira de Bioquímica e Biologia Molecular, 1996. 142p.        [ Links ]

12 COSTA H., PALMA MS. Agelotoxin: a phospholipase A2 from the venom of the neotropical social wasp cassununga (Agelaia pallipes pallipes) (Hymenoptera-Vespidae). Toxicon, 2000, 38, 1367-79.        [ Links ]

13 DESCHAUX O., BIZOT JC., GOYFFON M. Effects of apamin on learning and memory in rats. Toxicon, 1996, 34, 1087.        [ Links ]

14 DEVAUX C., KNIBIEHLER M., DEFENDINI ML., MABROUK K., ROCHAT H., VAN RIETSCHOTEN J., BATY D., GRANIER C. C-terminal amidation of apamin is important for biological activity as revealed by recombinant technology and chemical synthesis. Toxicon, 1996, 34, 1087.        [ Links ]

15 DOTIMAS EM., HIDER RC. Honeybee venom. Bee world, 1987,68, 51-71.         [ Links ]

16 DOTIMAS EM., HAMID KR., HIDER RC., RAGNARSON U. Isolation and structure analysis of bee venom mast cell degranulating peptide. Biochim. Biophys. Acta, 1987, 911, 285-93.        [ Links ]

17 EDSTROM A. Venomous and poisonous animals. Malabar: Krieger Publishing Company, 1992. 210p.        [ Links ]

18 FUNDENBERG HH., STITES DP., CAALDWELL JL., WELLS JV. Basic Clinical Immunology. Los Altos: Lange Med, 1980. 380p.        [ Links ]

19 GMACHL M., KREIL G. The precursor of the bee venom constituents apamin and MCD peptide are encoded by two genes in a tandem which share the same 3’ – exon. J. Biol. Chem., 1995, 270, 12704-8.        [ Links ]

20 GOLDEN DBK. Epidemiology of allergy to insect venoms and stings. Allergy Proc., 1989, 10, 103-7.        [ Links ]

21 GUTIÉRREZ JM., LOMONTE B. Local tissue damage induced by Bothrops snake venoms - A review. Mem. Inst. Butantan, 1989, 5, 211-23.        [ Links ]

22 HABERMANN E. Bee and wasp venoms. Science, 1972, 177, 314-22.        [ Links ]

23 HAIM B., RIMON A., ISHAY JS., RIMON S. Purification, characterization and anticoagulant activity of a proteolytic enzyme from Vespa orientalis venom. Toxicon, 1999, 37, 825-9.        [ Links ]

24 HANH G., LEDITSCHKE H. Apud BANKS BEC., SHIPOLINI RA. Chemistry and pharmacology of honey bee venom. In: PIEK T. Venoms of the Hymenoptera: Biochemical, pharmacological and behavioral aspects. London: Academy Press, 1986: 329-416.        [ Links ]

25 HEGNER D. Apud HABERMANN E. Bee and wasp venoms. Science, 1972, 177, 314-22.        [ Links ]

26 HIDER RC. Honeybee venom: a rich source of pharmacologically active peptides. Endeavour, 1988, 12, 60-5.        [ Links ]

27 HIDER RC., RAGNARSON U. A comparative study of apamin and related bee venom peptides. Biochim. Biophys. Acta, 1981, 667, 197-208.        [ Links ]

28 HOFFMAN DR. Hymenoptera venom proteins. Nat. Toxins, 1996, 2, 169-86.        [ Links ]

29 HOFFMAN DR., JACOBSON RS. Allergens in hymenoptera venom XII: how much protein is in a sting? Ann. Allergy, 1984, 52, 276-8.        [ Links ]

30 HOFFMAN DR., JACOBSON RS. Allergens in hymenoptera venom XXVII: bumblebee venom allergy and allergens. J. Allergy Clin. Immunol., 1996, 97, 812-21.        [ Links ]

31 KAMIGUTI AS., HANADA S. Study of the coagulant and proteolytic activities of newborn Bothrops jararaca venom. Toxicon, 1985, 23, 580.        [ Links ]

32 KEMENY DM., DALTON N., LAWRENCE J., PEARCE FL., VERNON CA. The purification and characterisation of hyaluronidase from the venom of the honeybee, Apis mellifera. Eur. J. Biochem., 1984, 139, 217-23.        [ Links ]

33 KOH Y., CHUNG K., KIM D. Biochemical characterization of a thrombin-like enzyme and a fibrinolytic serine protease from snake (Agkistrodon saxatilis) venom. Toxicon, 2001, 39, 555-60.        [ Links ]

34 KREIL G., BACHMAYER H. Biosynthesis of melittin, a toxic peptide from bee venom - detection of a possible precursor. Eur. J. Biochem., 1971, 20, 344.        [ Links ]

35 KUBELKA V., ALTMANN F., MÄRZ L. The asparagine-linked carbohydrate of honeybee venom hyaluronidase. Glycoconj. J., 1995, 12, 77-83.        [ Links ]

36 LIMA PRM., BROCHETTO-BRAGA MR. Caracterização de serina-proteases em venenos de diferentes subespécies de Apis mellifera (Hymenoptera - Apidae). In: CONGRESSO DE INICIAÇÃO CIENTÍFICA DA UNESP, 6, Guaratinguetá, 1994. Anais... Guaratinguetá: Universidade Estadual Paulista, 1994. 135.        [ Links ]

37 LIMA PRM., BROCHETTO-BRAGA MR., CHAUD-NETO J. Proteolytic activity of africanized honeybee (Apis mellifera: hymenoptera, Apidae) venom. J. Venom. Anim. Toxins, 2000, 6, 64-76. (SciELO)        [ Links ]

38 LOMONTE B., GUTIÉRREZ JM. La actividad proteolítica de los venenos de serpientes de Costa Rica sobre la caseína. Rev. Biol. Trop., 1983, 31, 37-40.        [ Links ]

39 MACKESSY SP. Characterization of the major metalloprotease isolated from the venom of the Northern Pacific rattlesnake, Crotalus viridis oreganus. Toxicon, 1996, 34, 1277-85.        [ Links ]

40 MUELLER U., REISMAN R., WYPYCH J., ELLIOT W., STEGER R., WALSH S., ARBESMAN C. Comparison of vespid venoms collected by electrostimulation and by venom sac extraction. J. Allergy Clin. Immunol., 1981, 68, 254-61.        [ Links ]

41 NABIL ZI., HUSSEIN AA., ZALAT SM., RAKHA MK. Mechanism of action of honey bee (Apis mellifera L.) venom on different types of muscles. Hum. Exp. Toxicol., 1998, 7, 185-90.        [ Links ]

42 NAKAJIMA T. Pharmacological biochemistry of vespid venoms. In: PIEK T. Venoms of the hymenoptera: biochemical, pharmacological and behavioral aspects. London: Academy Press, 1986: 309-24.        [ Links ]

43 NEUMAN W., HABERMANN E., AMEND G. Apud BANKS BEC., SHIPOLINI RA. Chemistry and pharmacology of honey bee venom. In: PIEK T. Venoms of the hymenoptera: biochemical, pharmacological and behavioral aspects. London: Academy Press, 1986: 329-416.        [ Links ]

44 OGAWA T., NAKASHIMA KI., NOBUHISA I., DESHIMARU M., SHIMOHIGASHI Y., FUKUMAKI Y., SAKAKI Y., HATTORI S., OHNO M. Accelerated evolution of snake venom phospholipase A2 isozymes to acquire diverse functions. Toxicon, 1996, 34, 287.        [ Links ]

45 OKANO M., NISHIZAKI K., SATOSKAR AR., YOSHINO T., MASUDA Y., HARN JR DA. Involvement of carbohydrate on phospholipase A2, a bee-venom allergen, in in vivo antigen-specific IgE synthesis in mice. Eur. J. Allergy Clin. Immunol., 1999, 54, 811-8.        [ Links ]

46 OWEN MD., PFAFF LA. Melittin synthesis in the venom system of the honey bee (Apis mellifera L.). Toxicon, 1995, 33, 1181-8.        [ Links ]

47 PATTANAARGSON S., ROBOZ J. Determination of hyaluronidase activity in venoms using capillary electrophoresis. Toxicon, 1996, 34, 1107-17.        [ Links ]

48 PIEK T. Insect venoms and toxins. In: KERKUT GA., GILBERT LI. Comprehensive insect physiology, biochemistry and pharmacology. Great Britain: Pergamon Press, 1985: 595-634.        [ Links ]

49 PRZYBILLA B. Bee and wasp allergy - clinical picture and diagnosis. J. Eur. Acad. Dermatol. Venereol., 1999, 9, S84.        [ Links ]

50 RUBERTI M., LIMA PRM., BROCHETTO-BRAGA MR. Comparative study on the protein composition and proteolytic activity of Hymenoptera venom. In: REUNIÃO ANUAL DA SOCIEDADE BRASILEIRA DE BIOQUÍMICA E BIOLOGIA MOLECULAR (SBBq), 28, Caxambú, 1999. Anais... Caxambú: Sociedade Brasileira de Bioquímica e Biologia Molecular, 1999. 19.        [ Links ]

51 SCHMIDT JO. Biochemistry of insect venoms. Ann. Rev. Entomol., 1982, 27, 339-68.        [ Links ]

52 SCHMIDT JO., BLUM MA., OVERALL WL. Comparative lethality of venom from stinging Hymenoptera. Toxicon, 1980, 18, 469-75.        [ Links ]

53 SCHMIDT JO., BLUM MS., OVERALL WL. Comparative enzymology of venoms from stinging hymenoptera. Toxicon, 1986, 24, 907-21.        [ Links ]

54 SHKENDEROV S. A protease inhibitor in bee venom. Identification, partial purification and some properties. FEBS Lett., 1973, 33, 343-7.        [ Links ]

55 SOUSA JRF., MONTEIRO RQ., CASTRO HC., ZINGALI RB. Proteolytic action of Bothrops jararaca venom upon its own constituents. Toxicon, 2001, 39, 787-92.        [ Links ]

56 TU AT., BAKER B., WONGVIBULSIN S., WILLIS T. Biochemical characterization of atroxase and nucleotide sequence encoding the fibrinolytic enzyme. Toxicon, 1996, 34, 295-300.        [ Links ]

57 WATALA C., KOWALCZYK JK. Hemolytic potency and phospholipase activity of some bee and wasp venoms. Comp. Biochem. Physiol., 1990, 97C, 187-94.        [ Links ]

58 WHAN U., THIEMEIER N., GENS C., FORCK G., KEMENY DM. The allergenic activity of purified bee venom proteins and peptides. J. Allergy Clin. Immunol., 1984, 73, 189.         [ Links ]

 

 

Correspondence to
M. R. Brochetto-Braga
Departamento de Biologia do Instituto de Biociências
Caixa Postal 199, 13.506-900 Rio Claro, São Paulo, Brasil
Email: mrbbraga@rc.unesp.br

Received June 26, 2002
Accepted June 28, 2002

Creative Commons License All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License