solubilization purifi cation and characterization of integralmembrane proteins

... 2005 FEBS Recognition of proteins containing CP2-binding motifs and verification of the CP2-binding ability of REST and YY1 that contain the HXPR motif As stated above, none of the 12-mer peptide ... The identification and characterization of human Sister -of- Mammalian Grainyhead (SOM) expands the grainyhead-like family of developmental transcription factors... & Andersen B (2003) ... more rounds of amplification as above. The phage titer of every round was determined by infecting 200 lL of a mid-log phase culture of E. coli with 10 lLof serially diluted phage and plating on

Ngày tải lên: 16/03/2014, 18:20

... studies of Mosimann et al [12], the hydroxyl group of Tyr179 lies within hydrogen-bonding distance of O7 and O9 of the glyceryl moiety of Neu5Ac, and a network of noncovalent bonds between enzyme and ... apparent values of kcatand Km(Neu5Ac), at a fixed CTP concentration of 154 lm and five different Neu5Ac concentrations, were 560 ± 30 s)1 and 190 ± 20 lm, respectively, in the presence of 0.2 mm dithiothreitol, ... pocket plays a role in substrate discrimination against binding of the 5-OH group of Kdo and in favour of the N-acetyl group of Neu5Ac and, although these residues are conserved in CNS enzymes, none

Ngày tải lên: 22/03/2014, 21:21

... a number of samples from serum stored over longer periods and fresh serum and found no evidence of the appearance of additional bands after storage (data not shown) Fig 3 Isolation of TNX from ... polyclonal antibody raised against FNIII27–28 of TNX This suggests that the N-terminus of the 75 kDa A B C D Fig 1 Identification and characterization of TNX species in serum. (A) The antigens to ... against FNIII29–30 of TNX and the signal for the bands corresponding to the 75 and 140– 150 kDa species are approximately of equal strength Most likely, the FNIII29–30 repeats of TNX are not as

Ngày tải lên: 23/03/2014, 09:21

... Trang 1Purification and characterization of three isoforms of chrysophsin,a novel antimicrobial peptide in the gills of the red sea bream, Chrysophrys major Noriaki ... melittin and more hemolytic than magainin (Fig 10) Fig 5 Cation-exchange HPLC of native P1, P2-2 and P3, and their synthetic peptides Mixtures of synthetic P1-COOH and P1-CONH 2 (A), native P1 and ... · 137.1 Fig 4 Cation-exchange HPLC of P1, P2 and P3 Aliquots of P1 (A), P2 (B) and P3 (C) were loaded on a PolySulfoethyl Aspartamide col-umn and eluted with a linear gradient of KCl in 5 m

Ngày tải lên: 23/03/2014, 20:22

... bu€er NiA and NiB; s, 0.2% CHS in solubilization bu€er and 0.02% in bu€ers NiA and NiB The quanti®cation of interactions with speci®c ligands allows the structural integrity and homogeneity of a puri®ed ... expression, solubilization and puri®cation of a hA2aR fusion protein in quantity and quality suf®cient for biophysical characterization and crystallization The following points made the puri®cation of ... Puri®cation (fold) 84 ± 762 1243 Ĩ FEBS 2002 Puri®cation and characterization of A2a receptor (Eur J Biochem 269) 87 Fig Puri®cation of hA2aR (M-A2aTr316-H10) from E coli cells The puri®cation

Ngày tải lên: 24/03/2014, 00:21

... consists of 137 amino acids (expected molecular mass of 15.266 kDa) and has a calculated pK value of 8.32 Gene cloning and protein purification The lys gene was cloned into plasmid vector pET3a, and ... Isolation and purification of endolysin Cells of E coli BL21(DE3) from 200 mL of culture (1.1 g) carrying plasmid pT5lys were suspended in 10 mL of 25 mm Tris⁄ HCl (pH 8.0), containing 40 mm NaCl and ... cell lysis and provide a competitive advantage to the phage under unfavorable growth conditions (analogs of the products of the Rz and Rz1 genes of bacterio-phage k) [36] One of these proteins

Ngày tải lên: 30/03/2014, 02:20

... expression and purification; Refolding of proteins Reagents and kits used; Bacterial strains and vectors used; Columns used for purification; Cloning of synthetic gene into pET-32a and pET-M; ... purification and cleavage of the fusion protein; RP-HPLC of untagged drysdalin; Refolding and RP-HPLC of untagged drysdalin; Expression of the His-drysdalin in pET-M; Affinity Purification of ... of venom and venom gland; RNA isolation and cDNA synthesis; Cloning of ds cDNA; Isolation of plasmids and verification of clones; cDNA sequencing; Sequence analysis; 3’RACE of venom proteins;

Ngày tải lên: 11/09/2015, 10:02

... Identification of the initiation codons in GmPDIL-3a and Gm-PDIL-3b mRNAs (A) Schematic representation of the structure of GmPDIL-3a and GmPDIL-3b mRNAs The putative ORFs (gray boxes) of GmPDIL-3a and ... ER proteins, or regulation of ER calcium channel activity [9–12] In plants, a set of 22 orthologs of known PDI family proteins was discovered by a genome-wide search of Arabidopsis thaliana, and ... N-glycosylation and the for-mation of disulfide bonds [1] Disulfide bonds are essential for structural stabilization and for regulation of the functions of many secretory and plasma mem-brane proteins

Ngày tải lên: 18/02/2014, 11:20

... GmPDIL-2 and proglycinin or b-conglycinin during the course of the folding process of these proteins Results Cloning and expression of GmPDIL-1 and GmPDIL-2 To clone the soybean ortholog of Arabidopsis ... region between b and b¢ in GmPDIL-1, and between b and b¢ and b¢ and a¢ in GmPDIL-2, the structure of these regions may be protease resistant The activity of recombinant GmPDIL-1 and GmP-DIL-2, ... cDNAs of GmPDIL-1 and GmPDIL-2 as members of the PDI family The amino acid sequences and putative domain structures of GmPDIL-1 and GmPDIL-2 were similar to each other, and both recombinant proteins

Ngày tải lên: 07/03/2014, 05:20

... Biology and Diabetes Research Centre, University of Linko¨ping, Sweden 2 Department of Molecular and Clinical Medicine, University of Linko¨ping, Sweden 3 Department of Medicine and Care and Diabetes ... 1 and 4, VHD-caveolae; lanes 2 and 5, HD-caveolae; lanes 3 and 6, LD-caveolae) The amount of the analysed proteins is expressed as arbitrary densitometric units and normalized to percentage of ... 9corroborated by the identification of the fatty acid-binding membrane proteins FATP1 and FATP4 in this specific class of caveolae FATP1 and FATP4 are the only FATP proteins expressed in adipocytes

Ngày tải lên: 16/03/2014, 13:20

... unfolding of the proteins is required for dissociation of the complex and membrane binding In the presence of membrane mimetics, the a-helical contents of Rv3619c and Rv3620 increased by 42% and 35%, ... Rv3618 and Rv3623 encode a probable monooxygenase and lipoprotein, respectively Rv3619c and Rv3620c are secretory proteins of 94 and 98 amino acids, respectively, reported in culture filtrates of ... for ESAT-6– CFP-10 and Rv0287–Rv0288 could result from the dif-ference in the folding state of Rv3620c from that of CFP-10 and Rv0287 Thermal unfolding of proteins The presence of any stable tertiary

Ngày tải lên: 22/03/2014, 16:21

... appearance, and biochemical properties Further characterization of both proteins and identification of specific interaction partners will contribute to a fuller understanding of the functions of thioredoxin ... Expression and purification of recombinant thioredoxin h Expression vectors of recombinant barley thioredoxin h isoforms were constructed for production of nontagged full-length thioredoxin h isoforms ... nmolÆs)1Ænmol)1and 13 lM Again, HvTrxh2 and TrxTa resembled each other with Vmax of 2.0 and 1.5 nmolÆs)1Ænmol)1and Kmof 44 lMand 37 lM, respect-ively In the same assay, a Kmvalue of 8 lMwas obtained

Ngày tải lên: 23/03/2014, 17:22

... activity of the recombinant proteins A large number of syncytia were observed when Assembly of VLPs by expression of RVFV N and Gc proteins Figure 5 Assembly of VLPs by expression of RVFV N and Gc proteins ... may facilitate understanding of these processes are highly desirable In this report we present the expression and characterization of RVFV structural proteins N, Gn and Gc and demonstrate the efficient ... complex structures To determine the posi-Expression and purification of RVFV nucleoprotein (N) protein Figure 1 Expression and purification of RVFV nucleoprotein (N) protein Insect Sf9 cells were

Ngày tải lên: 20/06/2014, 01:20

... 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

... 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

... 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