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1.
L. Zavalova S. Lukyanov I. Baskova E. Snezhkov S. Akopov S. Berezhnoy E. Bogdanova E. Barsova E. D. Sverdlov 《Molecular & general genetics : MGG》1996,253(1-2):20-25
We previously detected in salivary gland secretions of the medicinal leech (Hirudo medicinalis) a novel enzymatic activity, endo-ɛ(γ-Glu)-Lys isopeptidase, which cleaves isopeptide bonds formed by transglutaminase (Factor
XIIIa) between glutamine γ-carboxamide and the ɛ-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.
Received: 9 April 1996 / Accepted: 17 May 1996 相似文献
2.
3.
Pavel Bobrovsky Valentin Manuvera Izolda Baskova Svetlana Nemirova Alexandr Medvedev Vassili Lazarev 《Current issues in molecular biology》2021,43(3):2068
Leeches are amazing animals that can be classified as conditionally poisonous animals since the salivary cocktail they produce is injected directly into the victim, and its components have strictly defined biological purposes, such as preventing blood clot formation. Thrombolytic drugs are mainly aimed at treating newly formed blood clots. Aged clots are stabilized by a large number of isopeptide bonds that prevent the action of thrombolytics. These bonds are destroyed by destabilase, an enzyme of the leech’s salivary glands. Here, we conducted a pilot study to evaluate the feasibility and effectiveness of the use of destabilase in relation to blood clots formed during real pathological processes. We evaluated the isopeptidase activity of destabilase during the formation of a stabilized fibrin clot. We showed that destabilase does not affect the internal and external coagulation cascades. We calculated the dose–response curve and tested the ability of destabilase to destroy isopeptide bonds in natural blood clots. The effect of aged and fresh clots dissolving ability after treatment with destabilase coincided with the morphological characteristics of clots during surgery. Thus, recombinant destabilase can be considered as a potential drug for the treatment of aged clots, which are difficult to treat with known thrombolytics. 相似文献
4.
The medicinal leech Hirudo medicinalis produces various types of proteinase inhibitors: bdellins (inhibitors of trypsin, plasmin, and acrosin), hirustasin (inhibitor of tissue kallikrein, trypsin, -chymotrypsin, and granulocyte cathepsin G), tryptase inhibitor, eglins (inhibitors of -chymotrypsin, subtilisin, and chymasin and the granulocyte proteinases elastase and cathepsin G), inhibitor of factor Xa, hirudin (thrombin inhibitor), inhibitor of carboxypeptidase, and inhibitor of complement component C1s. This review summarizes data on their primary and tertiary structures, action mechanisms, and biological activities. 相似文献
5.
Tushmalova NA Pragina LL Baskova IP Basanova AV Zavalova LL 《Zhurnal vysshe? nervno? deiatelnosti imeni I P Pavlova》2001,51(2):252-253
In the paper are presented the results of the study of Pyavit, a complex leech preparation from, whose effects on the memory may be dependent on DNA methylation (one of the mechanisms of gene expression regulation). A positive effect of Pyavit on formation and retention of passive avoidance conditioned reaction was demonstrated in rat experiments. 相似文献
6.
Chalisova NI Pennijajnen VP Baskova IP Zavalova LL Bazanova AV 《Rossi?skii fiziologicheski? zhurnal imeni I.M. Sechenova / Rossi?skaia akademiia nauk》2001,87(12):1650-1655
Components of leech (Hirudo medicinalis) salivary gland secretion (high molecular mass bdelin B, bdellastasin, eglin C, destabilize) were used to investigate their neurotrophic effect on organotypic culture of dorsal root ganglia of the 10-11-day old chicken embryos. Destabilize and high molecular mass bdelin B in concentrations of 0.01, 0.02, 0.05 and 0.1 ng/ml, bdellastasin in concentrations of 0.02 and 0.05 ng/ml, eglin C in concentrations of 0.1 ng/ml exhibit neurite-stimulating effect as compared to the control. 相似文献
7.
I. P. Baskova T. G. Yudina L. L. Zavalova A. S. Dudkina 《Biochemistry. Biokhimii?a》2010,75(5):585-589
The relative location of proteins and lipids in particles of medicinal leech salivary gland secretion (SGS) is revealed for
the first time. Their sizes and morphology are described. Using scanning electron microscopy and transmission electron microscopy,
it was determined that SGS consists of particles of different sizes and form. This picture is supported by confocal laser
scanning microscopy of SGS preparations treated with fluorescein isothiocyanate. After incubation with nonionic detergents
(Brij 35 and Tween 20), transmission electron microscopy revealed the dissociation of fragments composing protein-lipid particles
(PLP), and in this case an increase in free protein concentration determined by a modification of the Lowry method was observed.
Perylene probing of lipids in SGS preparations showed that they are concentrated mainly inside PLP and are almost absent on
the surface. Cholesterol was detected during SGS probing using the cholesteryl-Bodipy (hydrophobic fluorescent analog of cholesterol)
on surface sections during confocal analysis of electron microphotographs of SGS. This analysis detected PLP structures in
SGS resembling caveoles full of cholesterol. SGS, preliminary frozen at −70°C, transformed into a multitude of similar small
particles visualized by transmission electron microscopy, whose fixed distribution resembled water crystal structure. 相似文献
8.
9.
Effects of common atopy-associated amino acid substitutions in the IL-4 receptor alpha chain on IL-4 induced phenotypes 总被引:3,自引:1,他引:3
Franjkovic I Gessner A König I Kissel K Bohnert A Hartung A Ohly A Ziegler A Hackstein H Bein G 《Immunogenetics》2005,56(11):808-817
The human IL-4 receptor alpha chain gene (IL4R) is highly polymorphic and controversial reports have been published with respect to the association of different single nucleotide polymorphisms (SNPs) with atopy markers. Here we analyzed the functional and associational relevance of common IL4R coding SNPs. Transfection of B cell lines expressing the IL-4R variant V75+R576 did not result in enhanced IL-4 induced CD23 expression compared to cell lines expressing the wild type IL-4R alpha chain. Transfection of the IL-4R variant P503 into a murine T cell line did not influence IL-4 induced T-cell proliferation compared to wild type constructs. Analysis of six IL4R coding SNPs (I75V, E400A, C431R, S436L, S503P, Q576R) and common haplotypes (frequency 0.05%) in blood donors (n=300) did not indicate a significant association with elevated serum IgE level. Moreover, the most informative IL4R coding SNPs (I75V, C431R, Q576R) and related two- and three-point haplotypes (frequency 0.05%) were analyzed in a second, extended group of blood donors (n=689). Again, no significant association with elevated serum IgE was detectable. We conclude that common coding SNPs in the IL4R gene are unlikely to contribute significantly to increased IgE levels and variations outside the coding region may influence atopy susceptibility. 相似文献
10.
Baskova IP Zavalova LL Basanova AV Moshkovskii SA Zgoda VG 《Biochemistry. Biokhimii?a》2004,69(7):770-775
Protein diversity of the high molecular weight fraction (molecular mass > 500 daltons) of salivary grand secretion of the medicinal leech Hirudo medicinalis has been demonstrated using methods of proteomic analysis. One-dimensional (1D) electrophoresis revealed the presence of more than 60 bands corresponding to molecular masses ranging from 11 to 483 kD. 2D-electrophoresis revealed more than 100 specific protein spots differing in molecular masses and pI values. SELDI-mass spectrometry analysis using the ProteinChip. System based on chromatography surfaces of strong anion or weak cation exchanger detected 45 individual compounds of molecular masses ranged from 1.964 to 66.5 kD. Comparison of SELDI-MS data with protein databases revealed eight known proteins from the medicinal leech. Other masses detected by proteomic analytical methods may be related to both modifications of known proteins and unknown biologically active components of leech saliva secretion. 相似文献