Hyaluronidase venom

A number of allergens from both honey bee and vespid venoms have been cloned and expressed by either Escherichia coli or baculovirus-infected insect cells (table 1) phospholipase Aj [20], hyaluronidase [21], acid phosphatase [13] and Api m6 [14] from honey bee venom, as well as antigen 5 [22], phospholipase A and hyaluronidase [23] from vespid venom, and dipeptidylpeptidases from both bee and Vespula venoms [15, 16]. Their reactivity with human-specific IgE antibodies to the respective allergens has been documented [11-16, 22, 23] and their specificity is superior... 

The toxicities of insect venoms are low to most people. Despite this, relatively large numbers of fatalities occur each year from insect stings because of allergic reactions in sensitized individuals. These reactions can lead to potentially fatal anaphylactic shock, which affects the nervous system, cardiovascular function, and respiratory function. The agents in bee venom that are responsible for severe allergic reactions are mellitin and two enzymes of high molecular mass — hyaluronidase and phospholipase A-2. 

Flavonoids are well known as inhibitors of hyaluronidase. Li 1950 Rodney et al, reported the effect of several flavonoids on hyaluronidase using in vitro and in vivo methods [79], Knppusamy et al. investigated the structure-activity relationship of die flavonoids as inhibitors of testicular hyaluronidase [80] and hyaluronidase from snake, scorpion, and bee venom [81]. They showed that flavones. flavanols. and chalcones possess the general ability to inhibit hyaluronidase Kinetic investigations showed that these compounds acted as competitive inhibitors [80]. Several hyaluronidase inhibitors were shown to be able to delay the action of snake venom when injected into mice [8ll One caimnt rule out the possible interaction of the flavonoids with other components of the venom, as the inhibitory effects of flavonoids are not specific to hyaluronidase... 

A. T. Tu and R. R. Hendon. Characterization of timid venom hyaluronidase and evidence for its action as a spreading factor. Comp. Biochem. PhyxioL 765 377 (1983). 

C. H. Puli, R. Yuen, M. C. M. Chung, and H. Khoo. Purification and partial characterization of hyaluronidase from stonetish Synanceja korrida) venom. Comp. Biachan. Physiol 70/0 159 (1992). 

P. G. Ricbman and H. Baer. A convenient plate asaay for the quantitation of hyaluronidase in hymenoptoa venoms. Anal Biockem. 109 376 (1960). 

Marz, L., Kuhne, C., and Michl, H. (1983). The glycoprotein nature of phospholipase As hyaluronidase and acid phosphatase from honey-bee venom. Tmdcan 21, 893— 896. 

Hyaluronidase activity is present in the venoms of a surprisingly wide variety of organisms. These include bees, wasps, hornets, spiders, scorpions, as well as certain species of fish, snakes, and lizards [203,204]. Some of these venoms, particularly those from snakes, have stretches of sequence with 36% identity with testicular hyaluronidase, PH-20. Interestingly, these hyaluronidases have absolute specificity for HA [205]. Hyaluronidase in venoms may act to facilitate penetration of other venom active ingredients. However, other evolutionary selective advantages of venom hyaluronidases may exist that have not yet been identified. 

Loxosceles venom contains hyaluronidase, alkaline phosphatase, 5-ribonucleotide phosphohydro-lase, and sphingomyelinase D. Sphingomyelinase D is a component of the cytotoxic venom, with a MW of... 

In humans, the specific disposition of venom is not well understood. The local distribution of venom is enhanced by the presence of hyaluronidase and other spreading factors found in the venom. Systemic absorption of venom components is likely dependent on lymphatic transport. The onset of local symptoms such as redness and pain may develop within a few hours of the bite. 

Brown Recluse spider venom contains many diverse protein fractions including spreading factors and enzymes such as hyaluronidase, collagenase, protease, phospholipase, and others. These venom components cause coagulation of blood and, ultimately, the occlusion of small blood vessels at the bite site. This leads to local skin and tissue necrosis due to ischemia. Hemolysis of red blood cells may also occur. The normal inflammatory processes that follow, such as edema and hemorrhage, contribute to the tissue damage caused by the venom. 

Animal venoms usually possess hyaluronidase activity [17]. The enzymatic properties, including hyaluronidase, of snake venoms have been extensively studied by Tan et al. [18]. Snake hyaluronidase acts on hyaluronan, chondroitin, and chondroitin-4- and -6-sulfate, producing various oligosaccharides, mainly tetrasaccharides [1]. 

Hyaluronidase from bee venom has about the same substrate specificity as snake hyaluronidase [19]. Venom of social wasps was found to contain high levels of hyaluronidase activity, whereas venom from ants contains low levels of activity [20]. Lizard venom contains a hyaluronidase that acts almost specific on hyaluronan, i.e., it has no activity toward chondroitin-6-sulfate, dermatan sulfate, or heparin and only weak activity toward chondroitin-4-sulfate [21]. 

A hyaluronidase has been purified and characterized from stonefish (Synanceja horrida) venom. It acts specifically on hyaluronan, producing tetra-, hexa-, octa-, and decasaccharides, but does not act on chondroitin sulfate or dermatan sulfate [22,23]. 

High amounts of hyaluronidase are present in scorpion venom [17]. The enzyme is also present in the venom of tarantula (Eurypelma califomicum) [24] and in the venom of the poisonous brown recluse spider (Loxosceles reclusa) [25],... 

Table 1 presents a survey of hyaluronidase activity in the venom of several animal species. Transglycosylation properties have been observed with hyaluronidase from snake venom (Crotalus terrificus) [8], but not with hyaluronidase from bee venom [19]. 

Table 1 Hyaluronidase Activities of Some Animal Venoms...

Hyaluronidases are generally glycoproteins. This has been confirmed for testicular [67], Staphylococcus aureus [68], lysosomal [59], lizard venom [21], and stonefish venom [22] hyaluronidases. The amino acid compositions of testicular [67], Staphylococcus aureus [68], lizard venom [21], and stone-fish venom [22] hyaluronidases have been determined. Of the latter type, the N-terminal sequence has been derived. The hyaluronidase gene of Streptococcus pyogenes bacteriophage H4489A has been expressed in Escherichia coli and the full nucleotide sequence has been derived. This made possible a mapping of the full amino acid sequence of the enzyme [69]. 

Figure 3 Inhibition properties of some flavonoids (250 mM) on hyaluronidase from several venom sources and from bovine testes. (From Refs. 80,81.)...

For this assay plates or petri dishes consisting of hyaluronan dispersed in agar and buffered to the appropriate pH are used. Cylindrical holes are punched in the gel and the samples (unknowns or standards) are added. The plates are incubated at 37°C for several hours. After incubation, a solution of a quaternary ammonium compound is poured over the gel, and after reaction the areas of digested hyaluronan become visible as clear rings. This technique is a simple assay for hyaluronidase activity in biological samples and has been used to estimate hyaluronidase activity in hymenoptera venoms [143], in semen, and in seminal plasma [146],... 

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