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A Phenylnorstatine Inhibitor Binding to HIV-1 Protease: Geometry, Protonation and Subsite-Pocket Interactions Analyzed at Atomic Resolution
Brynda, Jiří; Řezáčová, Pavlína; Fábry, Milan; Hořejší, Magdalena; Štouračová, Renata; Sedláček, Juraj; Souček, Milan; Hradilek, Martin; Lepšík, Martin; Konvalinka, Jan
2004 - anglický
The x-ray structure of a complex of HIV-1 protease (PR) with a phenylnorstatine inhibitor Z-Pns-Phe-Glu-Glu-NH2 has been determined at 1.03 A, the highest resolution so far reported for any HIV PR complex. The inhibiot shows subnanomolar Ki values for both the wild-type PR and the variant representing one of the most common mutations linked to resistance development. The structure displays a unique pattern of hydrogen bonding to the two catalytic aspartate residues. The high resolution permints to assess the donor/acceptor relations of this hydrogen bonding and to indicate a proton shared by the two catalytic residues. Structural mechanism for the unimpaired inhibition of the protease Val82Ala mutant is also suggested, based on energy calculations and analyses Byla určena rentgenová struktura komplexu proteázy HIV-1 (PR) s fenylnorstatinovým inhibitorem Z-Pns-Phe-Glu-Glu-NH(2) při rozlišení 1,03 A, nejvyšším rozlišení, které bylo doposud uvedeno pro kterýkoli komplex HIV PR. Tento inhibitor vykazuje subnanomolární hodnoty K(i) jak pro PR divokého typu, tak pro variantu představující jednu z nejběžnějších mutací spojenou s vývojem rezistence. Struktura obsahující fenylnorstatinovou část chirality (2R,3S) vykazuje jedinečný obraz vazby vodíkových můstků na dva katalytické aspartátové zbytky. Toto vysoké rozlišení umožňuje zjistit donorové a akceptorové vztahy těchto vodíkových můstků a určit proton sdílený oběma katalytickými zbytky. Je také navržen strukturální mechanismus nenarušené inhibice mutanty proteázy Val82Ala založený na energetických výpočtech a analýzách
Klíčová slova:
inhibitor; HIV protease; atomic resolution
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
A Phenylnorstatine Inhibitor Binding to HIV-1 Protease: Geometry, Protonation and Subsite-Pocket Interactions Analyzed at Atomic Resolution
The x-ray structure of a complex of HIV-1 protease (PR) with a phenylnorstatine inhibitor Z-Pns-Phe-Glu-Glu-NH2 has been determined at 1.03 A, the highest resolution so far reported for any HIV PR ...
Characterization of the clones derived from the HT29 colorectal cancer cell line
Šloncová, Eva; Kučerová, Dana; Vojtěchová, Martina; Turečková, Jolana; Tuháčková, Zdena; Sovová, Vlasta
2004 - anglický
During our analysis of HT29 cell line we established several clones, which differed in the expression of some molecular markers, e.g. phosphorylation of Src protein, PKCbeta2 activity, CEA-CAM1 expression and in the response to the differentiating effect of sodium butyrate, indicated by activity of alkaline phosphatase Během analýzy buněčné linie HT29 jsme připravili několik stabilních klonů, které se lišily expresí některých molekulárních markerů, např. fosforylací proteinu Src, aktivitou PKCbeta2, expresí CEA-CAM1 a odpovědí na diferenciační účinek butyrátu sodného
Klíčová slova:
colorectal carcinoma cells; differentiation
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Characterization of the clones derived from the HT29 colorectal cancer cell line
During our analysis of HT29 cell line we established several clones, which differed in the expression of some molecular markers, e.g. phosphorylation of Src protein, PKCbeta2 activity, CEA-CAM1 ...
Estimation of the post-meiotic state of spermatogenesis in man by immunofluorescence with monoclonal antibodies against intra-acrosomal sperm proteins
Chládek, David; Pěknicová, Jana
2003 - anglický
Male infertility due to severe azoospermia has been associated with the absence of spermatozoa in ejaculated. We tested the ejaculates (IVF latoratory Pranatal Ltd) from 30 patients with azoospermia for the presence of cells from defferent stages of spermiogenesis. Monoclonal sntibodews against intraacrosomal human sperm proteins were used in the immunofluorescence test for detection of acrosomal proteins in the acrosomal precursors and acrosomes.
Klíčová slova:
monoclonal antibody; acrosome
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Estimation of the post-meiotic state of spermatogenesis in man by immunofluorescence with monoclonal antibodies against intra-acrosomal sperm proteins
Male infertility due to severe azoospermia has been associated with the absence of spermatozoa in ejaculated. We tested the ejaculates (IVF latoratory Pranatal Ltd) from 30 patients with azoospermia ...
Dynamic and role of cytosekeletal proteins in mammalian sperm cells
Paleček, J.; Dvořáková, K.; Pěknicová, Jana; Kubátová, Alena; Dráber, Pavel
2003 - anglický
The cytoskeleton of sperm head consists of resistant structural proteins of the nucleus and of perinuclear theca, which are largely rosponsible for the shaping of the nucleus. three mahjor cyteskeletal proteins, actin, tzubnulin and spectrin, are present in the head of mammalian spermatohzoa. Maoreober, the mictotubular cytoskeleton also contains other dynamic proteins, which have a morphological function within the cell and which are responsible for cell signalling processes.
Klíčová slova:
cyprinus; carpio; acipenser
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Dynamic and role of cytosekeletal proteins in mammalian sperm cells
The cytoskeleton of sperm head consists of resistant structural proteins of the nucleus and of perinuclear theca, which are largely rosponsible for the shaping of the nucleus. three mahjor ...
Molecular cloning, E.coli expression and purification of SCFV antibody fragments of diagnostic/therapeutic interest
Král, Vlastimil; Fábry, Milan; Hořejší, Magdalena; Závada, Jan; Sedláček, Juraj
2003 - anglický
We describe molecular cloning, expression, purification and properties of two single chain antibody variable fragments (scFv) of potential diagnostic use, namely scFv M75 and scFv Tu-20. The former scFv is derived from a monoclonal antibody M75 specific for a cell surface protein MN/CA IX, strongly associated with many types of human carcinomas. The latter scFv is derived from a monoclonal antibody TU-20 specific for neuronal beta-III-tubulin.
Klíčová slova:
cloning; purification; scFv
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Molecular cloning, E.coli expression and purification of SCFV antibody fragments of diagnostic/therapeutic interest
We describe molecular cloning, expression, purification and properties of two single chain antibody variable fragments (scFv) of potential diagnostic use, namely scFv M75 and scFv Tu-20. The former ...
Characterization of peptide proteinase inhibitors isolated from boar seminal plasma
Jelínková, Petra; Tichá, M.; Jonáková, Věra
2003 - anglický
Most of proteins of boar seminal plasma are present in this medium under physiological conditions in the form of aggregates and not of monomers. In the present communication we have shown that not only spermadhesins, but also proteinase inhibitors participate in the formation of associated complexes. Proteinase inhibitors were found after gel chromatographic separation mainly in the fraction with the lowest relative molecular mass. Two different types of serine proteinase inhibitors were isolated from boar seminal plasma ů (i) glycosylated inhibitor with Mr 12 kDa, (ii) non-glycosylated sperm-associated acrosin inhibitor with Mr 8 kDa, and a new type of low molecular weight protein that is probably composed of two subunits coupled by the -S-S- bridge. The proteinase inhibitor with Mr 8 kDa was proved to form a heterodimer with AQN 1 spermadhesin under physiological conditions, whereas inhibitor with Mr 12 kDa forms homodimers.
Klíčová slova:
proteinase inhibitors, aggregated forms, boar seminal plasma
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Characterization of peptide proteinase inhibitors isolated from boar seminal plasma
Most of proteins of boar seminal plasma are present in this medium under physiological conditions in the form of aggregates and not of monomers. In the present communication we have shown that not ...
Antitrophoblast cell-mediated immunity /AT-CMI/ in recurrent miscarriage and ectopic pregnacy
Madar, J.; Kučera, E.; Šůla, K.; Chaloupková, A.; Pěknicová, Jana; Tolarová, M.; Nouza, K.; Kinský, R.
2003 - anglický
Specific mechanisms operate in decidua nad trophoblast to establish and maintain immunological tolerance of the embryo in mother's uterus. due to the lack of this tolerance, disturbances of pregnancy may result. In this presentation, we have tested the hypothesis that anti-trohhoblast cell-mediated immunity (AT-CMI) is elevated in habitually aborting women and that extra-uterine localisation of the embryo leads to subsequent elevation of AT-CMI.
Klíčová slova:
trophoblast; cardiolipin
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Antitrophoblast cell-mediated immunity /AT-CMI/ in recurrent miscarriage and ectopic pregnacy
Specific mechanisms operate in decidua nad trophoblast to establish and maintain immunological tolerance of the embryo in mother's uterus. due to the lack of this tolerance, disturbances of pregnancy ...
Multigenerational study - effect of the selected compounds on the mouse fertility
Boubelík, Michael; Pěknicová, Jana; Kyselová, Vendula; Buckiová, D.
2003 - anglický
Several compounds known to be effective for their influence onto the environment were tested. Compounds tested in this study were: benzo/a/ pyrene, lindane (hexachlorocyclohexane isomer), 4-n-nonylphenol, bisphenol A and octachlorostyrene. The question was if these compounds might also function as endocrine disruptors.
Klíčová slova:
endocrine disruptors; sperm motility
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Multigenerational study - effect of the selected compounds on the mouse fertility
Several compounds known to be effective for their influence onto the environment were tested. Compounds tested in this study were: benzo/a/ pyrene, lindane (hexachlorocyclohexane isomer), ...
Microsatellite region in the murine oncogene src - its primary structure and variability during evolution
Fučík, Vladimír; Šindelka, Radek; Beran, Jaroslav; Ferjentsik, Zoltán; Jonák, Jiří
2003 - anglický
In the fifth intron of the Mus musculus oncogene src a complex microsatellite was discovered that consists of a CA run and 31 bp apart followed by track of TG dinucleotides interrupted by extra Gs in specific intervals. As the 31 bp interlink can assume a hairpin structure, the whole region can be viewed as an imperfect inverted repeat. The microsatellite was sequenced in imbred lines of M. m. musculus and M. m. domesticus, as well as in mice of both subspecies trapped near the hybrid zone. Differences in the length of repeat regions, insertion/deletions and simple base changes characterize the strains. Hybrids between the two species could be identified by sequencing of cloned PCR products. The structure of the microsatellite in more distant species was found to be conspicuously different. Instead of frequent (CA)18-25, we can see only (CA)8 in M. spretus, CATGCGCCCCCCA in Mus pahari and CACCCCCC in Mus caroli. No similar structure is present in the corresponding human src intron.
Klíčová slova:
Mus musculus, c-src introns, microsatellite
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Microsatellite region in the murine oncogene src - its primary structure and variability during evolution
In the fifth intron of the Mus musculus oncogene src a complex microsatellite was discovered that consists of a CA run and 31 bp apart followed by track of TG dinucleotides interrupted by extra Gs in ...
A Phenylnorstatine Inhibitor Binding to HIV-1 Protease: Geometry, Protonation and Subsite-Pocket Interactions Analyzed at Atomic Resolution
Brynda, Jiří; Řezáčová, Pavlína; Fábry, Milan; Hořejší, Magdalena; Štouračová, Renata; Sedláček, Juraj; Souček, M.; Hradílek, M.; Lepšík, M.; Konvalinka, J.
2003 - anglický
The x-ray structure of a complex of HIV-1 protease (PR) with a phenylnorstatine inhibitor Z-Pns-Phe-Glu-Glu-NH2 has been determined at 1.03 A, the highest resolution so far reported for any HIV PR complex. The inhibiot shows subnanomolar Ki values for both the wild-type PR and the variant representing one of the most common mutations linked to resistance developoment. The structure displays a unique pattern of hydrogen bonding to the two catalytic aspartate residues. The high resolution permits to assess the donor/acceptor realtions of this hydrogen bonding and to indicate a proton shared by the two catalytic residues. Structural mechanism for the unimpaired ihnibition of the protease Val82Ala mutant is also suggested, based on energy calculations and analyses.
Klíčová slova:
inhibitor; HIV protease; atomic resolution
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
A Phenylnorstatine Inhibitor Binding to HIV-1 Protease: Geometry, Protonation and Subsite-Pocket Interactions Analyzed at Atomic Resolution
The x-ray structure of a complex of HIV-1 protease (PR) with a phenylnorstatine inhibitor Z-Pns-Phe-Glu-Glu-NH2 has been determined at 1.03 A, the highest resolution so far reported for any HIV PR ...
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