Monomerization of fragment dd by destabilase from the medicinal leech does not alter the N-terminal sequence of the γ-chain

Zavalova, L. L.; Kuzina, E.V.; Levina, N.B.; Baskova, I. P.

doi:10.1016/0049-3848(93)90099-a

Cited by 19 publications

(8 citation statements)

References 3 publications

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“…1. To detect fractions with isopeptidase activity and to characterise the purified enzyme we assayed specific hydrolysis of -( -Glu)-Lys isopeptide bonds between chains of the so-called D-D fibrin dimer, a product of partial hydrolysis of stabilized fibrin (Zavalova et al 1993) (Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

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Genes from the medicinal leech (Hirudo medicinalis) coding for unusual enzymes that specifically cleave endo-ɛ(γ-Glu)-Lys isopeptide bonds and help to dissolve blood clots

et al. 1996

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We previously detected in salivary gland secretions of the medicinal leech (Hirudo medicinalis) a novel enzymatic activity, endo-epsilon(gamma-Glu)-Lys isopeptidase, which cleaves isopeptide bonds formed by transglutaminase (Factor XIIIa) between glutamine gamma-carboxamide and the epsilon-amino group of lysine. Such isopeptide bonds, either within or between protein polypeptide chains are formed in many biological processes. However, before we started our work no enzymes were known to be capable of specifically splitting isopeptide bonds in proteins. The isopeptidase activity we detected was specific for isopeptide bonds. The enzyme was termed destabilase. Here we report the first purification of destabilase, part of its amino acid sequence isolation and sequencing of two related cDNAs derived from the gene family that encodes destabilase proteins, and the detection of isopeptidase activity encoded by one of these cDNAs cloned in a baculovirus expression vector. The deduced mature protein products of these cDNAs contain 115 and 116 amino acid residues, including 14 highly conserved Cys residues, and are formed from precursors containing specific leader peptides. No homologous sequences were found in public databases.

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Section: Resultsmentioning

confidence: 99%

“…The enzyme specifically splits isopeptide bonds in diisopeptide (Baskova et al 1990), and between fibrin chains, but leaves polypeptide chains intact (Zavalova et al 1991(Zavalova et al , 1993. It dissolves stabilized fibrin in vitro (Baskova and Nikonov 1985) and stimulates thrombolysis in experimental animals .…”

Section: Introductionmentioning

confidence: 99%

Genes from the medicinal leech (Hirudo medicinalis) coding for unusual enzymes that specifically cleave endo-ɛ(γ-Glu)-Lys isopeptide bonds and help to dissolve blood clots

et al. 1996

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“…Destabilase acts as a thrombolytic agent in leech; it cleaves peptide bonds and promotes brinolysis, thereby inhibiting coagulation activity [23][24][25][26]. As a multifunctional enzyme, destabilase can also exert antibacterial function.…”

Section: Destabilase-related Genesmentioning

confidence: 99%

Transcriptome Assembly of Asian Buffalo Leech Hirudinaria Manillensis and An Accurate Identification of Genes Involved in Anticoagulant Biosynthesis

Fan¹,

Abbood²,

Lu³

et al. 2020

Preprint

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Background: The Asian buffalo leech Hirudinaria manillensis is a valuable animal widely used in traditional Chinese medicine to cure blood-clotting. However, although the content of the anticoagulant genes in H. manillensis were identified, their actual expression profile remains undetermined. Herein, in this study we conducted transcriptome analysis on this species to address this subject.Results: A high qualified accurate gene expression profile of H. manillensis transcripts was obtained. By identifying genes upregulated during feeding, anticoagulant-related genes and signaling pathways were identified. The assembled Unigenes were compared in various mainstream databases, and a total of 16,682 Unigenes received annotation information. In addition, the Unigenes were evaluated in terms of length distribution, GC content, and expression level. The data showed good sequencing quality and high reliability. A total of 155 anticoagulant-related genes were found in this study, including those involved in different types of degradation of the extracellular matrix and inhibition of platelet aggregation.Conclusions：Substantial transcriptome information was obtained by transcriptome sequencing of H. manillensis. This information should help to provide a further molecular theoretical basis for functional gene analysis, genomics, genetic diversity analysis, and molecular marker development of H. manillensis and for the anticoagulation-related genes of this species and its medicinal value as an anticoagulant.

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“…The isopeptidase activity of destabilase involves the hydrolysis of the stabilized fibrin and its proteolytic degradation product D-dimer [1,6]. Hydrolysis occurs not by common proteolytic cleavage but by disruption of the e-(c-Glu)-Lys isopeptide bonds between polypeptide chains [7]. In an experiment using animal models, it was found that this mechanism of action results in the slow disruption of preformed thrombi, i.e., destabilase is a thrombolytic agent.…”

Section: Introductionmentioning

confidence: 98%

Generation of recombinant destabilase-lysozyme from medicinal leeches in three different expression systems

Manuvera

Kurdyumov

Filonova

et al. 2015

Protein Expression and Purification

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Monomerization of fragment dd by destabilase from the medicinal leech does not alter the N-terminal sequence of the γ-chain

Cited by 19 publications

References 3 publications

Genes from the medicinal leech (Hirudo medicinalis) coding for unusual enzymes that specifically cleave endo-ɛ(γ-Glu)-Lys isopeptide bonds and help to dissolve blood clots

Genes from the medicinal leech (Hirudo medicinalis) coding for unusual enzymes that specifically cleave endo-ɛ(γ-Glu)-Lys isopeptide bonds and help to dissolve blood clots

Transcriptome Assembly of Asian Buffalo Leech Hirudinaria Manillensis and An Accurate Identification of Genes Involved in Anticoagulant Biosynthesis

Generation of recombinant destabilase-lysozyme from medicinal leeches in three different expression systems

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