solubilization purifi cation and characterization of integralmembrane proteins

... tissue of mouse and spleen and thymus of human and mouse [11,37] Foxp3 is also highly expressed in lymphoid organs, such as the thymus and spleen [49], and its expression within a variety of other ... t-bet, stat6 and foxp3 were also determined and found to be very similar to their human homologues Both the human and zebrafish T-bet and Foxp3 genes contained the same number of exons and introns ... the expression of these genes within humans and mice, of T-bet within Ginbuna crucian carp [27] and STAT6 in mandarin fish [28] have shown similar widespread expression In the case of STAT6, highest...

Ngày tải lên: 16/02/2014, 09:20

... properties of SmSmad1 and SmSmad1B as described above However, in the presence of SmSmad4 and the active mutant form of TbRI, both SmSmad2 and SmSmad4 interacted with GCN5, indicating the formation of ... from S mansoni Fig Structure of the SmSmad1B gene, cDNA and protein (A) Schematic representations of the SmSmad1B gene, cDNA and protein and the amino acid sequence of SmSmad1B protein Five exons ... location was determined to be the position of the putative transcription start site (Fig 1B) Analysis of the 5¢ upstream region of exon demonstrated the lack of a conserved TATA box upstream of...

Ngày tải lên: 18/02/2014, 16:20

... This paper reports the puri cation and characterization of GAT from wild watermelon leaves, and discusses its function during drought ⁄ strong-light stress on the basis of its two unique enzymatic ... experiments and vertical bars are SD (B) Immunoblot analysis of DRIP-1 in the total soluble proteins (20 lg per lane) isolated from plants before (0 day) and after 1, 2, and days of drought ⁄ ... (DTT) and 20% (v ⁄ v) glycerol, and 20% saturation of (NH4)2SO4 The enzyme was eluted with a linear gradient of 30–0% saturation of (NH4)2SO4 in buffer A Active fractions were collected and applied...

Ngày tải lên: 20/02/2014, 03:20

... De-Oacetylation of glycoproteins was performed following the procedures of Sarris and Palade [30] and Schauer [6] A solution of 750 lL of glycoprotein (5 mgÆmL)1) was added to 250 lL of 0.04 M of NaOH, vortexed, ... well and mixed The values of HAI titer were determined after 60 of incubation and expressed as the highest dilution of glycoprotein that inhibited the agglutination of erythrocytes Sialoglycoproteins ... revealed that puri cation of lectin from the hemolymph of crustaceans was most successful by affinity chromatography as it gave a higher fold of puri cation and percentage of recovery [20,24,27,35–39]...

Ngày tải lên: 21/02/2014, 00:20

... released orthophosphate was measured by a modi cation of the method of Martin and Doty [21], using toluene instead of benzene To keep background hydrolysis of the acid-labile phosphoamidate as low ... absence of mM magnesium chloride inhibit the enzyme Puri cation of the recombinant phosphohistidine phosphatase Puri cation of phosphohistidine phosphatase from porcine liver A 9-mL sample of the ... cascade Pharmacol Ther 67, 323–350 Wei, Y.F & Matthews, H.R (1991) Identi cation of phosphohistidine in proteins and puri cation of protein-histidine kinases Methods Enzymol 200, 388–414 Fothergill-Gilmore,...

Ngày tải lên: 21/02/2014, 01:21

... is the first report of the puri cation of a 4-amino3-hydroxybenzoate-fission enzyme and its characterization in terms of molecular mass, subunit structure, reaction mechanism and catalytic properties ... buffer A Proteins were eluted with a linear gradient (0–0.4 M) of NaCl in 900 mL of buffer A Fractions of mL were collected at a flow rate 40 mLÆh)1 The protein concentration and enzyme activity of ... 1,2,4-trihydroxybenzene and pyrogallol) in aqueous solution were always freshly prepared and used immediately Effect of various compounds on the enzyme activity The effect of metal salts, and chelating and sulfhydryl...

Ngày tải lên: 21/02/2014, 01:21

... maximum of the a band at 557 nm absorption maxima at 420 nm (c band), 530 nm (b band) and 557 nm (a band) are characteristic of cytochrome b Heme was extracted from the protein with acidic acetone and ... behavior is typical of integral membrane proteins The polypeptide with an apparent molecular mass of 53 kDa appears as a double band in unboiled samples (lanes A1 and B1) Identi cation of the genes ... Beifuss, U & Deppenmeier, U (1998) Isolation and characterization of methanophenazine and function of phenazines in membranebound electron transport of Methanosarcina mazei Go1 J Bac¨ teriol 180,...

Ngày tải lên: 21/02/2014, 03:20

... with GroEL from 15 and 30 lg of the cell lysate without (lanes and 3) or in the presence of ATP (lanes and 4) was detected with anti-UP12 Ig Samples (3 and lg of total proteins) of the cell extract ... Effect of addition of mM DNP on cell growth (h) and expression of UP12 (j) (B) Monitoring of cell growth (s) and amount of UP12 (d) before and after addition of 0.1 mM CCCP (C) Effect of temperature ... the following observations support the classi cation of UP12 as a member of the UspA family Sub-cloning, puri cation, and characterization of UP12 and its interaction with GroEL In order to investigate...

Ngày tải lên: 22/02/2014, 07:20

... min, bands with molecular masses of 43 and 39 kDa were observed in addition to undigested bands of ZP2 and ZP3 After incubation for 10 min, three major bands with molecular masses of 43, 39 and ... phylogenetic analysis of fish hatching enzyme genes, hatching enzyme originally consisted of a single type of enzyme, and HCE and LCE were produced by duplication and diversi cation of the gene [16] ... mechanism of its egg envelope digestion and compared it with that of medaka hatching enzyme Finally, the evolution of the mechanism of egg envelope digestion is discussed on the basis of the manner of...

Ngày tải lên: 07/03/2014, 04:20

... regulation of a number of genes [7–10] Study of SmRXR partners will add to our knowledge of nuclear receptor gene regulation in schistosomes and to an understanding of the evolution of RXR’s function ... Isolation and characterization of Schistosoma mansoni constitutive androstane receptor Mol Biochem Parasitol 148, 31–43 14 Lu C, Wu W, Niles EG & LoVerde PT (2006) Identi cation and characterization of ... development by iQ5 application (cDNA is diluted to ·1-, ·10-, ·100- and ·1000-fold; see protocol of Bio-Rad iQ5 application) Normalized gene expression [23] of SmNR1 was calculated and standardized to...

Ngày tải lên: 07/03/2014, 11:20

... Fractionation and puri cation of the enzymes stored in the latex of Carica papaya J Chrom B 790, 229–238 Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage ... acid sequences of the proteins VXH-I, VXH-III, VXH-IVa and VXH-IVb are shown in Fig 6, and are compared with the known N-terminal sequences of the cysteine proteinases of C papaya and V cundinamarcensis ... latex of Carica candamarcensis Hook Biol Chem Hoppe-Seyler 374, 501–506 Walraevens V, Vandermeers-Piret M, Vandermeers A, Gourlet P & Robberecht P (1999) Isolation and characterization of the...

Ngày tải lên: 07/03/2014, 11:20

... transcriptional start site, and the BRE motif, targets for the general transcription factors TBP and TFB, respectively [17] bcp2 mRNA B 16S rRNA Puri cation and characterization of recombinant Bcp2 (rBcp2) ... range of concentrations (0– 100 lgÆmL)1) (B) Effect on protection against DNA cleavage of the replacement of Cys49 of rBcp2 with serine Lane 1, pUC19; lane 2, pUC19 and 10 mM DTT; lane 3, pUC19 and ... absence of putative catalases and the presence of four putative Bcps proteins: Bcp1, Bcp2, Bcp3, Bcp4 whose roles should be clarified in detail and could play a key role in the detoxi cation processes...

Ngày tải lên: 07/03/2014, 12:20

... and Xenopus lysates, which recognises the PR65 a and b isoforms equally well The best anti-C monoclonal was found to be F2 6A10, and it recognizes PP2AC in rabbit, mouse and Xenopus oocytes and ... The full-length cDNA of XPR130 and XN73 were then used in an in vitro transcription–translation reaction, and the products separated by SDS/PAGE Proteins of about 130 kDa and 50 kDa, respectively, ... revealed that a large part of the 5¢-UTR and the complete N-terminal Ó FEBS 2003 specific part of HPR130 reside in one exon of 2435 bp (exon 2a) Comparison of the XN73 and XPR130 sequences with...

Ngày tải lên: 08/03/2014, 08:20

... Influences of temperature and pH on enzyme activity For the determination of the optimal temperature of the enzyme, the reaction was carried out at 25, 30, 35, 40, 45, 50, 55, 60, 65 and 70 °C and the ... amount of D-amino acids produced was determined by the Chirobiotic T HPLC method and the appropriate D-amino acids were used as the standards RESULTS Identi cation and phylogenetic analysis of the ... activity of N-D-AAase is affected by the presence of metal ions Thus, the enzyme was treated with EDTA, EGTA, 1,10-phenanthroline and metal ions at concentrations of and 10 mM The presence of the...

Ngày tải lên: 08/03/2014, 16:20

... N-Terminal sequencing of the kininogens was only successful from the 42-kDa band of wolffish kininogen (XLVQPGVLI…, Table 1) The major bands of both kininogens and also the 60-kDa band of cod kininogen ... kininogen and 6200 UÆmg)1 for wolffish kininogen (units are defined Fig SDS/PAGE visualized by silver staining of purified cod kininogen and wolf fish kininogen, and MALDI-TOF mass spectra of kininogens ... N-Terminus of fragment V N Ra d Glycopeptide Determination of partial amino-acid sequences Fig Inhibition of papain (0.625 mg) by different amounts of cod kininogen and wolffish kininogen as lg of papain...

Ngày tải lên: 08/03/2014, 23:20

... electrophoresis and N-terminal sequence of the purified enzyme (A) Native PAGE of the purified enzyme (0.5 lg) in the absence of SDS (B) SDS ⁄ PAGE of the purified enzyme (2 lg) and N-terminal sequence of bands ... fragments may be degradation products of CBCP produced in the course of the puri cation Characterization of daikon CBCP Isolation of cDNA encoding the catalytic domain of daikon radish CBCP In order ... 5435 Characterization of daikon CBCP A Tsuji et al A B C Fig Cleavage of glucagon, ACTH and b-casein by daikon CBCP HPLC profiles of glucagon (A) and ACTH (B) digests Peaks were collected and sequenced...

Ngày tải lên: 16/03/2014, 04:20

... [55] Expression and purication of the recombinant N-terminal region of FXII and domain D5 of HK Expression vectors for the recombinant N-terminal region of FXII and domain D5 of HK were constructed ... dtu.dk/services/SignalP/) Production and purication of the recombinant proteins Triafestin-1 and triafestin-2 recombinant proteins were produced in a baculovirusinsect cell system Triafestin-1 and triafestin-2 full-length ... Fig SDS PAGE and western blot analysis of a T infestans salivary gland extract and puried recombinant triafestin-1 and triafestin-2 A crude salivary gland extract (lane 1), lg of recombinant...

Ngày tải lên: 16/03/2014, 05:20

... L(small)-Trp as an endogenous ligand which binds to site II, and of cefazolin (site I-ligand) and verapamil (site I and II-ligand) as exogenous ligands All these ligands are known for their high ... towards l-Trp of HSA This might also be related to increased levels of serotonin and the frequent occurrence of some adverse complications of diseases which have an increased level of oxidized ... and not homocysteine, adducts The comparative FTMS measurement of peptide mixture derived from highly oxidized HSA standard (HSA-Cys) and purified HSA of healthy human plasma From the result of...

Ngày tải lên: 16/03/2014, 13:20

... reaction was stopped by the addition of 750 lL of acetic acid, and the color developed by the addition of 250 lL of ninhydrin (4 mgÆmL)1) at 95 °C for h A standard curve of different ornithine concentrations ... of sporulating microorganisms II The occurrence and partial puri cation of an arginase J Biol Chem 241, 620–623 26 Shimotohno, K.W., Iida, J., Takizawa, N., EndC, T (1994) ¸ Puri cation and characterization ... presence of cobalt, and then heat activating the Puri cation and catalytic activity of RocF expressed in E coli The rocF gene was translationally fused to the His6encoding construct, pQE30, and shown...

Ngày tải lên: 16/03/2014, 16:20

... Fig Kinetic characterization of r-Pfen (A) Plot of [2-PGA] vs activity; and (B) plot of [PEP] vs activity for the determination of Km A lL sample of enzyme containing 1.5 and 3.0 lg of r-Pfen, ... ampli cation of the enolase gene from the gametocyte cDNA library of the NF54 strain of E coli resulted in a fragment of the expected size of 1.4 kb This fragment was cloned in pQE30 vector, and ... response of r-Pfen to various concentrations of KCl is at variance to those of mammalian and yeast enolases Figure 5B shows the effect of increasing concentrations of Mg2+ on the activity of r-Pfen,...

Ngày tải lên: 16/03/2014, 18:20