Tài liệu Báo cáo Y học: Regulated expression and intracellular localization of cystatin F in human U937 cells pptx
Tài liệu Báo cáo Y học: Regulated expression and intracellular localization of cystatin F in human U937 cells pptx
... staining of cystatin F and ER.(A) Cystatin F was stained with a human cystatin F specific polyclonal antiserum and aFITC labelled secondary antibody (green sig-nal). (B) Following the cystatin F staining, ... cystatin F produced by U937 cells isretained intracellularly or is taken up by the cells immedi-ately following secretion. Accordingly, the relatively lowamount of cystatin F produced by U937 ... (cystatin C numbering).Recombinant cystatin F expressed in Sf9 insect cells isindeed glycosylated [5].Compared to other cystatins, cystatin F is an unusuallyspecific inhibitor. Among the family...
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Tài liệu Báo cáo khoa học: Tissue expression and biochemical characterization of human 2-amino 3-carboxymuconate 6-semialdehyde decarboxylase, a key enzyme in tryptophan catabolism pptx
... malaria and in the brain of a murine model of the syndrome[18,19]. Interestingly, picolinate is reportedly able toprevent the neurotoxic effects of quinolinate in the ratcentral nervous system, ... suc-cessfully restored the bacterial protein [21,33].Interestingly, the same difficulty in stripping off themetallocofactor from the protein has been describedfor murine adenosine deaminase, ... surroundingresidues involved in catalysis is more hydrophobic in the human, rather than in the bacterial protein.Finally, this study demonstrated the expression in kidney and liver of an alternatively...
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Tài liệu Báo cáo khoa học: Functional expression and mutational analysis of flavonol synthase from Citrus unshiu pptx
... substitution of glycine residues by alanine or proline and the exchange of proline by glycine. Glycine residues are frequently found atthe C-terminal ends of a helices providing the conforma-tional ... by site-directed mutagenesis of FHT [7,9] and by documentation of ligand binding in crystalline DAOCS and IPNS complexes [23–25]. Thecoordination of FeIIis commonly mediated by two histidine and ... Germany) asrequired for the substitution of glycine by alanine and proline, respectively, or of proline by glycine (Table 1). Eachindividual mutation was verified by DNA sequencing usingthe...
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Tài liệu Báo cáo khoa học: Complex transcriptional and translational regulation of iPLA2c resulting in multiple gene products containing dual competing sites for mitochondrial or peroxisomal localization docx
... pGL3-Enhancer(Promega) and then utilized for LipofectAMINE Plus-mediated transien t transfection of CV1 ce lls followed 24 hlater by analysis of luciferase activity utilizing the Lucife raseAssay System o f Promega. ... by heat-ing a 4-lMmixture of primers to 95 °C for 3 min followedby cooling to 22 °C prior to cloning into the Xho1/Sal1sites of vector pEGFP-N3. Integrity and orientation of theN-terminal ... witheach of the inhibitory constructs was performed usingLipofectAMINE Plus (Invitrogen). For each transfection,1–2 lg of luciferase r eporter plasmid was cotransfected with100 n g of pcDNA 3.1/myc-His/lacZ...
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Tài liệu Báo cáo khoa học: Endogenous expression and protein kinase A-dependent phosphorylation of the guanine nucleotide exchange factor Ras-GRF1 in human embryonic kidney 293 cells pptx
... kDa and 60 kDa isoforms of Ras-GRF1 (Fig. 2A, left panel) and of recombinant HA-Ras-GRF1 (Fig. 2B, upperpanel). Phosphorylation of ERK1 ⁄ 2 in the same sam-ples was fully induced after 3 min ... increasephosphorylation of endogenous Ras-GRF1 in ratbrain [9].Serotonin-induced phosphorylation of Ras-GRF1is dependent on cAMP and PKASerotonin increases adenylyl cyclase activity in HEK293 cells ... Allthree Raf isoforms [A-Raf, B-Raf and Raf-1 (C-Raf)]Keywords5-HT7, cAMP, ERK, GEF, serotoninCorrespondence F. O. Levy, Department of Pharmacology,University of Oslo, PO Box 1057 Blindern,N-0316...
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Tài liệu Báo cáo Y học: Ligand interactions and protein conformational changes of phosphopyridoxyl-labeled Escherichia coli phosphoenol pyruvate carboxykinase determined by fluorescence spectroscopy pdf
... that obtained in the presence of Mn2+(Table 1).An independent estimation of the binding affinity of PEPfor E. coli PEP carboxykinase was obtained from thequenching of the emission of Trp residues ... only in the presence of Mn2+. This high binding affinity of PEP in the presence of Mn2+appears common to PEP-carboxykinases. A Kd of 0.6 lMhas been determined forthe dissociation of PEP ... highbinding affinity of PEP in the presence of Mn2+[28]. Thehigh affinity of PEP in the presence of Mn2+suggests aspecific interaction between these two ligands in the enzymeactive site. In...
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Tài liệu Báo cáo Y học: The glucose-specific carrier of the Escherichia coli phosphotransferase system Synthesis of selective inhibitors and inactivation studies pptx
... reactivity of, Cys421 towards a second molecule.Alternatively, biphasic inactivation by 1a (with two inacti-vation rates differing by a factor of 10) may originate fromthe different accessibility of ... wererecently introduced for the preparation of C-6 aldehydederivatives of Glc [15]. The key step is the use of Collinsreagent for the selective oxidation of the primary trimethyl-silyl ether of the fully ... plotted in the Eadie–Hofsteeform (Fig. 2). In the absence of an inhibitor, EIIGlcdisplayed biphasic kinetics (Fig. 2, solid symbols), consis-tent with the presence of two binding sites of differentaffinity...
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Tài liệu Báo cáo Y học: Identification and characterization of a mammalian 14-kDa phosphohistidine phosphatase pdf
... was from FlukaChemie AG, Switzerland. Phenylmethanesulfonyl fluoride and okadaic acid were from Sigma. Trypsin of modifiedsequencing grade was obtained from Promega, and LysC of Achromobacter lytii ... liver cytosolFor small-scale preparation of cytosol, 10 g of freshporcine liver obtained from the local abattoir washomogenized for 1 min at 600 r.p.m. in three volumes of homogenization buffer ... h.Purification of phosphohistidine phosphatasefrom porcine liver cytosolA mixture of 320 mL 10 mMHepes, pH 7.5 and 320 mL of cytosol from 130 g of liver, was stirred for one hour with600 mL of DEAE-cellulose,...
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Tài liệu Báo cáo Y học: The RuvABC resolvasome Quantitative analysis of RuvA and RuvC assembly on junction DNA doc
... further evidence of the formation of the RuvAC-complex.Fig. 6. SPR sensorgram showing the DNA binding specificity of RuvA.(A) Binding of wild-type RuvA (2 lM) and (B) binding of the mutantRuvA ... different flow cells on astreptavidin sensor chip. The binding of 2 lME. coli RuvAwith the different complexes is shown in Fig. 6A. Thedifference in stability of the duplex and three-strand-RuvAcomplex ... nonspecific binding and instrument drift.RESULTSStoichiometry and kinetics of the RuvA–Hollidayjunction interactionThe stoichiometry and kinetics of the RuvA–Hollidayjunction interaction was analysed...
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Tài liệu Báo cáo Y học: Insights into the reaction mechanism of Escherichia coli agmatinase by site-directed mutagenesis and molecular modelling ppt
... cata-lyses the hydrolysis of agmatine to putrescine and urea [1].Agmatine, which results from decarboxylation of arginineby arginine decarboxylase [2], is a metabolic intermediate in the biosynthesis ... according to the instructions of the manufacturer. A single protein band was detected bySDS/PAGE of purified enzymes.Enzyme activities were determined by measuring theformation of urea from ... as shown in Fig. 2, it was half inactivated bydialysis against 10 mMEDTA in 5 mMTris/HCl (pH 7.5)for 4 h at 4 °C and full recovery of enzyme activity wasproduced by incubation of the EDTA-treated...
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