Structural analogues of bradykinin and other vasoactive peptides in the venom of the wasp Polistes gallicus

Six vasoactive peptides have been isolated in the homogeneous state from the venom of the waspPolistes gallicus. Their full amino acid sequences have been determined. It has been shown that four of them are structural analogues of bradykinin with additional N- and C-terminal sequences and intramolecular substitutions of amino acids. Some structure-function relationships in bradykinin analogues are discussed.A. S. Sadykov Institute of Bioorganic Chemistry, Uzbek SSR Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 389–393, May–June, 1990.     

Vasoactive peptides from venom of the wasp Polistes gallicus. Isolation and physicochemical and functional properties

Six vasoactive peptides have been isolated in the homogeneous form from the venom of the waspPolistes gallicus. Their physicochemical characteristics have been investigated. The wasp kinins exhibit myotropic and hypotensive effects. One of the kinins shows a prolonged hypotensive action. A. S. Sadykov Institute of Bioorganic Chemistry, Uzbek SSR Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 564–568, July–August, 1989.     

Vasoactive peptides from venom of the wasp Polistes gallicus. Isolation and physicochemical and functional properties

Six vasoactive peptides have been isolated in the homogeneous form from the venom of the waspPolistes gallicus. Their physicochemical characteristics have been investigated. The wasp kinins exhibit myotropic and hypotensive effects. One of the kinins shows a prolonged hypotensive action.A. S. Sadykov Institute of Bioorganic Chemistry, Uzbek SSR Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 564–568, July–August, 1989.     

Dominulin A and B: Two new antibacterial peptides identified on the cuticle and in the venom of the social paper wasp Polistes dominulus using MALDI-TOF, MALDI-TOF/TOF, and ESI-ion trap

Two new antibacterial peptides, denominated as Dominulin A and B, have been found on the cuticle and in the venom of females of the social paper wasp Polistes dominulus. The amino acidic sequence of the two peptides, determined by mass spectrometry, is INWKKIAE VGGKIL SSL for Dominulin A (MW = 1854 Da) and INWKKIAEIGKQVL SAL (MW = 1909 Da) for Dominulin B. Their presence on the cuticle was confirmed using MALDI-TOF by means of micro-extractions and direct analyses on body parts. The presence in the venom and the primary structure of the dominulins suggest their classification in the mastoparans, a class of peptides found in the venom of other Aculeate hymenoptera. Their antimicrobial action against Gram+ and Gram- bacteria fits in the range of the best natural antimicrobial peptides. Dominulins can represent an important defense of the colony of Polistes dominulus against microbial pathogens.     

Resistance of vasoactive peptides from venom of Vespa orientalis to action of proteolytic enzymes

The kinetics of the hydrolysis by proteolytic enzymes of vasoactive peptides isolated from the venom of the hornetVespa orientalis has been investigated. It has been found that the half-period of decomposition of the peptides exceeds that of bradykinin. These facts are in harmony with in vivo results on the prolonged hypotensive action of the peptides.A. S. Sadykov Institute of Bioorganic Chemistry, Uzbek SSR Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 5, pp. 658–661, September–October, 1988.     

Sequencing wasp venom peptides by endopeptidase digestion and nested collision-induced dissociation/post-source decay methods

A method incorporating nested collision-induced dissociation/post-source decay (CID/PSD) combined with endopeptidase digestion is described as an approach to determine the sequence of N-terminally modified peptides. The information from immonium and related ions observed in the CID/PSD spectrum was used for the selection of a suitable endopeptidase for the digestion of peptides. Rapid and reliable assignment of peptide sequence was performed by the comparison of CID/PSD spectra of both intact and endopeptidese-digested peptide fragments, since the assignments of the observed fragment ions to either N- or C-terminal ions can thus be carried out unambiguously. This nested CID/PSD method was applied to the sequence determination of two peptides from the solitary wasps Anoplius samariensis and Batozonellus maculifrons (pompilid wasps), which could not be sequenced by the Edman method due to N-terminal modification.     

Mass spectrometric characterization of two novel inflammatory peptides from the venom of the social wasp Polybia paulista

The social wasp P. paulista is relatively common in southeast Brazil causing many medically important stinging incidents. The seriousness of these incidents is dependent on the amount of venom inoculated by the wasps into the victims, and the characteristic envenomation symptoms are strongly dependent on the types of peptides present in the venom. In order to identify some of these naturally occurring peptides available in very low amounts, an analytical protocol was developed that uses a combination of reversed-phase and normal-phase high-performance liquid chromatography (HPLC) of wasp venom for peptide purification, with matrix-assisted laser desorption/ionization time-of-flight post-source decay mass spectrometry (MALDI-Tof-PSD-MS) and low-energy collision-induced dissociation (CID) in a quadrupole time-of-flight tandem mass spectrometry (QTof-MS/MS) instrument for peptide sequencing at the sub-picomole level. The distinction between Leu and Ile was achieved both by observing d-type fragment ions obtained under CID conditions and by comparison of retention times of the natural peptides and their synthetic counterparts (with different combinations of I and/or L at N- and C-terminal positions). To distinguish the isobaric residues K and Q, acetylation of peptides was followed by Q-Tof-MS analysis. The primary sequences obtained were INWLKLGKMVIDAL-NH(2) (MW 1611.98 Da) and IDWLKLGKMVMDVL-NH(2) (MW 1658.98 Da). Micro-scale bioassay protocols characterized both peptides as presenting potent hemolytic action, mast cell degranulation, and chemotaxis of polymorphonucleated leukocyte (PMNL) cells. The primary sequences and the bioassay results suggest that these toxins constitute members of a new sub-class of mastoparan toxins, directly involved in the occurrence of inflammatory processes after wasp stinging.     

Structural characterization of novel chemotactic and mastoparan peptides from the venom of the social wasp Agelaiapallipes pallipes by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry

High-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS) and high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) techniques were applied for the detection, purification, monitoring, and sequencing of two novel and biologically active peptides occurring at very low levels in the venom of the wasp Agelaia pallipes pallipes. These peptides were sequenced under LC/ESI-MS/MS conditions and designated as Agelaia-CP (I/L-L-G-T-I-L-G-L-L-K-G-I/L-NH2, MW 1207.8 Da) and Agelaia-MP (I/L-N-W-L-K-L-G-K-A-I-I-D-A-I/L-NH2, MW 1565.0 Da). The peptide Agelaia-CP showed no hemolytic activity, but it behaved as a mast cell degranulator and induced a potent chemotaxis in polymorphonucleated leukocyte (PMNL) cells, typical of a wasp chemotactic peptide. The peptide Agelaia-MP showed both powerful mast cell degranulation and hemolysis of washed rat red blood cells, and is thus assigned as a new member of the mastoparan family of peptides. Both peptides seem to be directly involved in the strong inflammatory reactions associated with wasp stings.     

Synthesis and biological properties of bradykinin-potentiating peptides from snake venom

Summary A pentapeptide and undecapeptide possessing bradykinin-potentiating action have been synthesized, and they have been subjected to comparative pharmacological investigation.M. M. Shemyakin Institute of Bioorganic Chemistry, Academy of Sciences of the USSR. Translated from Khimiya Prirodnykh Soedinenii, No. 1, pp. 47–56, January–February, 1975.     

Investigation of the cardiotoxic peptides of the venom of the cobraNaja oxiana

Six populations of basic polypeptides have been found in the venom of the Central Asian cobraNaja oxiana Eichwald, and four of them — V″c-1, V″c-2, V″c-5, and V″c-6 — have been obtained in the electrophoretically homogeneous state. On a perfused frog heart preparation, all the basic polypeptides proved to be cardiotoxic. However, only some of them (V′c-2, V″c-3, and V″c-4) were characterized by arrhythmic activity as well as by the negative ino- and chronotropic effects, common for all of them. An enhancement of the specific cardiotoxic effect of each of the polypeptides by pure phospholipase A-2 has been shown.     

Vasoactive peptides from venom of the hornet Vespa orientalis physiochemical and functional characterization

Two peptides having kinin activity have been isolated from hornet venom. The peptides isolated exhibit hypotensive action and a myotropic effect, and they also liberate histamine from mast cells. One of the peptides is a close structural analog of bradykinin while the other differs considerably from it.Institute of Bioorganic Chemistry of the Uzbek SSR Academy of Sciences, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 255–258, March–April, 1988.     

Inhibition of angiotensin-converting enzyme by peptides from the venom of the viperEchis multisquamatus

Angiotensin I-converting enzyme [dipeptidyl carboxypeptidase] (ACE, CE 3.4.15.1) isolated in the pure form from ox kidney, has been characterized physicochemically and kinetically. Inhibition of the enzyme by homogeneous peptides from the venom of the Central Asian viperEchis multisquamatus Ch. has been investigated. Institute of Biochemistry, Academy of Sciences of the Republic of Uzbekistan, Tashkent, fax (371) 62 32 56. Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 821–825, November–December, 1998.     

Separation and analysis of peptides and proteins from Vipera lebetina snake venom

Our studies of Levantine viper venom have demonstrated that the venom is a rich source of biomedically important proteins and peptides. The venom contains metalloproteases: thrombolytic, fibrin-degrading lebetase, an endothelial cell apoptosis inducing metalloprotease (VLAIP), factor X activator (VLFXA); serine proteases: factor V activator, bradykinin-releasing serine proteases, β-fibrinogenase, α-fibrinogenase and chymotrypsin-like protease and different other enzymes such as phosphodiesterase, 5`-nucleotidase, ribonuclease, phospholipase A2s and L-amino acid oxidase. Among nonenzymatic components venom contains: nerve growth factor, vascular endothelial growth factor, disintegrins, C-type lectins.Here we report the isolation and characterization of proteins and peptides from Vipera lebetina venom using size exclusion, ion exchange, hydrophobic interaction and affinity chromatography, HPLC, UPLC and MALDI-TOF MS methods. N-terminal sequences and internal sequences of tryptic peptides of different proteins were determined using Edman sequencing and LC-ESI-MS/MS techniques. On the basis of fragmental sequences of proteins the oligonucleotides were designed and used as primers for cDNA cloning. Using cDNA library of the venom gland of a single snake the cDNAs coding proteins were cloned and sequenced. Protein sequences were deduced from cDNA sequences.The substrate specificity of venom proteases against insulin B-chain, bradykinin, substance P, and 6-10 amino acid residues containing peptides synthesized according to potential cleavage regions of fibrinogen, factor X, factor IX, factor V, α₂-macroglobulin bait region and pregnancy zone protein were studied using MALDI-TOF mass spectrometry technique.     

Advantages of using nested collision induced dissociation/post-source decay with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: sequencing of novel peptides from wasp venom

We have examined the applicability of the 'nested' collision induced dissociation/post-source decay (CID/PSD) method to the sequencing of novel peptides from solitary wasps which have neurotoxic venom for paralyzing other insects. The CID/PSD spectrum of a ladder peptide derived from an exopeptidase digest was compared with that of the intact peptide. The mass peaks observed only in the CID/PSD spectrum of a ladder peptide were extracted as C-terminal fragment ions. Assignment of C-terminal fragment ions enabled calculation of N-terminal fragment masses, leading to differentiation between N-terminal fragment ions and internal fragment ions. This methodology allowed rapid and sensitive identification by removing ambiguity in the assignment of the fragment ions, and proved useful for sequencing unknown peptides, in particular those available as natural products with a limited supply.     

Investigation of the cardiotoxic peptides of the venom of the cobraNaja oxiana

Six populations of basic polypeptides have been found in the venom of the Central Asian cobraNaja oxiana Eichwald, and four of them — Vc-1, Vc-2, Vc-5, and Vc-6 — have been obtained in the electrophoretically homogeneous state. On a perfused frog heart preparation, all the basic polypeptides proved to be cardiotoxic. However, only some of them (Vc-2, Vc-3, and Vc-4) were characterized by arrhythmic activity as well as by the negative ino- and chronotropic effects, common for all of them. An enhancement of the specific cardiotoxic effect of each of the polypeptides by pure phospholipase A-2 has been shown.Institute of Biochemistry, Academy of Sciences of the Republic of Uzbekistan, Tashkent. Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 416–420, May–June, 1994.     

Methyl calcein blue and other analogues of calcein blue

4-Methylumbelliferone-8-methylenesarcosine (Methyl Calcein Blue) and four related metallofluorochromic indicators derived from umbelliferone, 4-methylumbelliferone, and 4-methylesculetin by condensation with formaldehyde and iminodiacetic acid or glycine have been synthesized, the structures established, the absorbance and fluorescence measured as functions of pH, and the reactions with copper(II) and calcium studied with attention to the effects on fluorescence. All of the compounds display a maximum fluorescence at ph about 9. The fluorescence of each is quenched by copper(II). The calcium derivatives of the compounds derived from the umbelliferones and iminodiacetic acid fluoresce at high ph but those from the umbelliferones and glycine or sarcosine do not. At high pH, 4-methylesculetin and the amino-acids derived from it do not fluoresce either alone or in the prescence of calcium.