Destabilase, the novel &amp;epsiv;-(??-Glu)-Lys isopeptidase with thrombolytic activity

Baskova, I. P.; Nikonov, G I

doi:10.1097/00001721-199102000-00025

Cited by 57 publications

(30 citation statements)

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“…For example, the saliva of H. medicinalis was found to inhibit plasma kallikrein and to contain destabilase [25], which liquefies cross-linked fibrin and is antithrombotic in rats [35]. Another compound called antistasin also has been identified in medicinal leeches.…”

Section: Bioactive Compounds In Leech Salivamentioning

confidence: 99%

An overview of leech and its therapeutic applications

2015

J Coast Life Med

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Section: Bioactive Compounds In Leech Salivamentioning

confidence: 99%

An overview of leech and its therapeutic applications

2015

J Coast Life Med

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“…191,192 Destabilase, a leech enzyme that hydrolyzes cross-links, provides an alternative approach to reversing the consequences of factor XIIIa-mediated fi brin crosslinking. 193,194 Neither of these agents has been tested in humans.…”

Section: Factor Xiiia Inhibitorsmentioning

confidence: 99%

New Antithrombotic Drugs

Weitz

Eikelboom

Samama

2012

Chest

284

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“…This enzyme activity leads to dissolution of stabilized fibrin. Destabilase was first described in the salivary glands of the leech Hirudo medicinalis , 55 later shown to be the product of a multigene family that is related to the lysozyme superfamily. 56,57 Five ESTs in the S. nigrimanum sialotranscriptome have the destabilase CDD motif, possibly coding for two alleles.…”

Section: Ubiquitous Protein Families Of Characterized Function(s)mentioning

confidence: 99%

An Insight into the Sialotranscriptome of Simulium nigrimanum, a Black Fly Associated with Fogo Selvagem in South America

Ribeiro

Valenzuela

Pham

et al. 2010

The American Society of Tropical Medicine and Hygiene

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Abstract. Pemphigus foliaceus is a life threatening skin disease that is associated with autoimmunity to desmoglein, a skin protein involved in the adhesion of keratinocytes. This disease is endemic in certain areas of South America, suggesting the mediation of environmental factors triggering autoimmunity. Among the possible environmental factors, exposure to bites of black flies, in particular Simulium nigrimanum has been suggested. In this work, we describe the sialotranscriptome of adult female S. nigrimanum flies. It reveals the complexity of the salivary potion of this insect, comprised by over 70 distinct genes within over 30 protein families, including several novel families, even when compared with the previously described sialotranscriptome of the autogenous black fly, S. vittatum. The uncovering of this sialotranscriptome provides a platform for testing pemphigus patient sera against recombinant salivary proteins from S. nigrimanum and for the discovery of novel pharmacologically active compounds.*Address correspondence to José M. C. Ribeiro, 12735 Twinbrook Parkway room 2E32D, Rocvkville, MD 20852. E-mail: jribeiro@niaid .nih.gov This paper should be a guide to browsing the supplemental files S1 and S2, found at: http://exon.niaid.nih.gov/transcriptome/S_nigrima num/S1/Sn-S1-web.xls ; http://exon.niaid.nih.gov/transcriptome/S_nig rimanum/S2/Sn-S2-web.xls . 1061SIMULIUM SIALOTRANSCRIPTOME strand. This sequence is then used in subsequent PCR reactions and restriction digests.First-strand synthesis was carried out using PowerScript reverse transcriptase at 42°C for 1 hour in the presence of the SMART IV and CDS III (3′) primers. Second-strand synthesis was performed using a long distance (LD) PCR-based protocol, using Advantage Taq polymerase (Clontech) mix in the presence of the 5′ PCR primer and the CDS III (3′) primer. The cDNA synthesis procedure resulted in creation of SfiI A and B restriction enzyme sites at the ends of the PCR products that are used for cloning into the phage vector (lambda TriplEx2 vector, Clontech). The PCR conditions were as follows: 95°C for 1 min; 24 cycles of 95°C for 10 sec, 68°C for 6 min. A small portion of the cDNA obtained by PCR was analyzed on a 1.1% agarose gel to check quality and range of cDNA synthesized. Double-stranded cDNA was immediately treated with proteinase K (0.8 μg/mL) at 45°C for 20 min, and the enzyme was removed by ultrafiltration though a Microcon (Amicon Inc., Beverly, CA) YM-100 centrifugal filter device. The cleaned, double-stranded cDNA was then digested with SfiI at 50°C for 2 hours, followed by size fractionation on a ChromaSpin-400 column (Clontech) into small (S), medium (M), and large (L) transcripts based on their electrophoresis profile on a 1.1% agarose gel. Selected fractions were pooled and concentrated using a Microcon YM-100.The concentrated cDNA mixture was ligated into the λ TriplEx2 vector (Clontech), and the resulting ligation mixture was packaged using the GigaPack III Plus packaging extract (Stratagene, La Jolla, CA) according ...

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Destabilase, the novel ϵ-(??-Glu)-Lys isopeptidase with thrombolytic activity

Cited by 57 publications

References 0 publications

An overview of leech and its therapeutic applications

An overview of leech and its therapeutic applications

New Antithrombotic Drugs

An Insight into the Sialotranscriptome of Simulium nigrimanum, a Black Fly Associated with Fogo Selvagem in South America

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