signaling autophagy and amino acid specificity

... and 450 nm Molecular mass information was obtained as described [6], determining the elution volumes of standard proteins and the void volume of the column The Superdex 200 PC 3.2/30 column and ... N210K and copurification of the 4¢-phosphopantothenoylCMP intermediate His-CoaB proteins (His-CoaB and His-CoaB N210K) were purified from E coli cell extracts by Ni-nitrilotriacetic acid and either ... (forward) 5¢-GCCAGTTTTGAATTCTGGAAAGCGC CTCG-3¢ and (reverse) 5¢-CGGGTCCAAGATCTTAA CGTCGATTTTTTTC-3¢ as primers (introduced EcoRI and BglII sites are underlined) and the purified chromosomal DNA as template...

Ngày tải lên: 19/02/2014, 12:20

... uncharged polar amino acids (i.e Q, N, S, and T) and an increase in charged amino- acid residues (i.e K, E, and R) in hyperthermophilic proteins As S and T can catalyze the deamination and backbone ... The amounts of fructose and xylulose produced were determined using the cysteine/carbazole/sulfuric acid method Absorbance was measured at 537 nm and 560 nm for xylulose and fructose, respectively ... 0.981, is based on the elution data of the 200, 150, and 66 kDa protein standards Linear regressions (1) and (2) give molecular masses of 200 and 177.5 kDa, respectively, for BLXI q FEBS 2001...

Ngày tải lên: 22/02/2014, 07:20

... barrel between the b3-strand and the a3-helix; (b) the catalytic machinery consists of the b4-strand aspartate (nucleophile), b5-strand glutamate (proton donor) and b7-strand aspartate (transition-state ... heavy-chain subunit of these amino acid transporters from the enzymes involved in the metabolism of starch and related saccharides Results and Discussion Evolutionary relationships and sequence–structural ... especially in higher animals (mammals and also in frogs and fishes), an aspartate (aspartic acid 248 in human 4F2hc; aspartic acid 380 in Fig as both the N-terminal and transmembrane segments are involved)...

Ngày tải lên: 18/02/2014, 13:20

... striking feature of conomarphin The l -amino acid to d -amino acid epimerization in a polypeptide chain is quite rare and not well understood, although some d -amino acids have been known for a long ... worth pointing out that the l -amino acid to d -amino acid epimerization seems to be complete for conopeptides, whereas both isoforms coexist in defensin-like peptides and natriuretic peptides from ... methods on an ABI 433A peptide synthesizer using standard Fmoc chemistry and side-chain protection N-terminal sequencing and MS N-terminal amino acid sequence analysis was performed by automated...

Ngày tải lên: 18/02/2014, 17:20

... contamination from xylem, and vice versa Glx (glutamine and glutamate: ve carbon amide and amino acid) and Asx (asparagine and aspartate: four carbon amide and amino acid) were found to be the ... Amino acid composition in leaves, and amino acid percentage ratio in the phloem exudates and leaves and xylem bleeding sap and leaves in the light and dark The amino acid composition in leaves is ... asparagine were the major amino acids in the xylem sap in both the light (63% of the total amino acids) and dark (60%) (Fig 7B) The amino acid percentage ratio in the leaves and phloem distinguished...

Ngày tải lên: 18/02/2014, 18:20

... W140O and K133A] The curves of W140A and W140O are nearly linear in terms of intensity All protein concentrations were mgÆmL)1 of the wild-type protein and K133A and E142O were 88.45, 84.70, and ... decrease in helicity on removal of all amino acids beyond position 140 In CD spectra in the near-UV region (Fig 1B), the wild-type protein and mutants E142O and K133A show strong intensity at  ... truncations and point mutations to C-terminal residues on SNase stability and conformation integrity was examined by subjecting these mutants and wild-type proteins to CD [11,12] and differential...

Ngày tải lên: 20/02/2014, 01:20

... stressed and the relaxed conformations [15] The network involves the side chains of the amino acids in positions 53 and 56 in a helix B, 186 in b strand 3A, and position 334 in b strand 5A (Fig ... b strand 6A and the backbone of Asn171 in the a helix F/b strand 3A loop Note: SWISSPDB VIEWER uses the same signature for a helices and the short 310-helix found in the a helix F/b strand 3A ... number of substitutions in the shutter region and b strand 5A of PAI-1 and studied their effect on the transition to latent and substrate forms and on the stabilizing effect of vitronectin, a...

Ngày tải lên: 22/02/2014, 07:20

... amino acid- regulated genes according to their biological processes reveals that amino acid limitation regulates groups of genes that are involved in amino acid and protein metabolism, lipid and ... components of the amino acid control of gene expression are not yet completely understood in mammals: (a) the genes and biological processes regulated by amino acid availability, and (b) the signaling ... to amino acid availability has been studied extensively in yeast [8] The GCN2 and TOR kinases sense the intracellular concentration of amino acids In addition, yeast cells possess an amino acid...

Ngày tải lên: 07/03/2014, 03:20

... yellow and the fruit is red? G Giorio et al Fig Sequence alignment of PSY1 and PSY2 protein derived from analysis with The predicted protein sequences were 412 and 438 amino acids for PSY1 and PSY2, ... and Psy2 genes and also provide the deduced complete amino acid sequences of the two enzymes, together with new insights into the transcriptional regulation of Psy1 and Psy2 in chloroplast- and ... photoprotection (quenching and xanthophylls cycle) and in the formation of abscisic acid Moreover, many species use them to make coloured flowers and fruits to attract pollinators and seed dispersers...

Ngày tải lên: 07/03/2014, 05:20

... chain glycine L-Alanine L-a-Aminobutyric acid L-Norvaline L-Norleucine Elongation of the backbone b-Alanine c-Aminobutyric acid d-Aminopentanoic acid e-Aminohexanoic acid O-Methyl substitution ... inward currents and affinity, d-aminopentanoate or e-aminohexanoate failed to induce any transport currents (Table 1) Methyl-substitutions at the amino- or carboxy-group of the a -amino acids had a ... spectrum that includes not only the amino acids glycine, alanine, serine and proline but also osmolytes such as sarcosine and betaine, and the D-enantiomers of serine and alanine The apparent affinities...

Ngày tải lên: 07/03/2014, 16:20

... with H121 and K9, and of G129 with K110 and K133 Other charged amino acids such as D73, D83 and E101 reinforce the interactions of E75 and E129 ˚ (OE1 ⁄ OE2)– are K110 (Nz)+ (¼ 6.29 A) and K133 ... Local stability and key acidic amino acids in staphylococcal nuclease by the interactions of E75 with H121 and K97 It is believed that a small number of key amino acid segments play major ... electrostatic bond between E75 and H121, and a hydrogen bond between D77 and T1208 Thus interactions between D77 and E75 and surrounding positively charged amino acids may result in the cooperative...

Ngày tải lên: 07/03/2014, 21:20

... three-dimensional structures of a -amino acids and those of b -amino acids and posed the following questions Which structures of b -amino acids overlap the canonical structures of a -amino acids? Subsequently, ... (Eur J Biochem 270) Fig Schematic representation of a-, b2- and b3 -amino acids with the torsion angles /, h and w For the b2 and b3 -amino acid substituted-SP analogues, the methyl side chain of b-HAla ... conformation of a -amino acids In order to visualize on the potential energy surfaces the conformers of b -amino acids that fit with canonical conformations of a -amino acids, two (/ ) w) Ramachandran maps...

Ngày tải lên: 08/03/2014, 08:20

... N-acyl-D- or L -amino acids and other D -amino acid derivates was determined (Table 3) The substrates analysed were a range of hydrophilic, hydrophobic and aromatic N-acyl or derivative D -amino acids that ... 20 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 Identification and phylogenetic analysis ... may prefer hydrophobic amino acids such as D-methionine and D-leucine N-acetyl derivates to aromatic amino acid such as D-phenylalanine and D-tryptophan N-acetyl derivates and that there is chiral...

Ngày tải lên: 08/03/2014, 16:20

... substituted Lys47 with three different amino acids, i.e acidic, neutral and basic amino acids The fact that eIF5A activity is impaired partially by Ala substitution and totally by Asp substitution, ... site-directed mutagenesis of each conserved amino acid, and by truncation, tested them as substrates for DHS and DOHH, and assessed their activities in supporting growth and protein synthesis in an eIF5A ... was no cross-reactivity between antibodies to human and yeast eIF5A (compare lanes 17 and 18 in Fig 2C, and lanes and in Fig 2D) The yeast eIF5A and most of the heIF5A mutants were readily detectable...

Ngày tải lên: 16/03/2014, 06:20

... CORE, the LID and the NMPbind [1] A characteristic of bacterial CMP kinases is an extension of the NMPbind domain by 40 amino acid residues forming a three-stranded antiparallel b sheet and two a ... structures of CMP and UMP, and interactions between the cytosine moiety of nucleotide and various side chains of wild-type CMP kinase (Upper) Chemical differences between CMP and UMP are indicated ... (D132S, D132N and D132H) led to a moderate decrease in stability, lowering Tm by 45 C compared with the wild-type protein The last group includes amino acid substitutions (D132A and R110M) that...

Ngày tải lên: 16/03/2014, 10:20

... confirmed to have a d -amino acid, and synthetic or expressed versions with both l -amino acids and d -amino acids at the correct locus in order to predict when and where d -amino acids may be found post-translationally ... correct amino- acid sequences of conotoxins r11b and r11c have d-Phe and d-Leu at positions 44 and 42, respectively Henceforth, we will refer to the d version of the peptides simply as r11b and r11c, ... modifies the l- amino acid to the d -amino acid does not stringently recognize a long vicinal sequence around the residue to be modified, nor is it specific for a particular amino- acid side chain...

Ngày tải lên: 16/03/2014, 22:20

... various steroid hormones (androsterone, epitestosterone) and fatty acids [7] UGT2B4 and UGT2B7 therefore play a key role in the detoxification of cholestatic bile acids and may prevent the formation ... UGT2B7Y33F and UGT2B4 and 200 lg for UGT2B4F33Y and UGT2B4F33L in 100 mm Tris ⁄ HCl buffer (pH 7.4), 10 mm MgCl2 containing 0.02 mm UDP-glucuronic acid and 0.1 lCi UDP-[U-14C]glucuronic acid The ... study was to identify amino acid residues that are involved in substrate specificity of UGTs 2B4 and 2B7 in order to better understand the molecular basis of substrate recognition and catalysis by...

Ngày tải lên: 23/03/2014, 09:21

... proteinogenic amino acids and C1 metabolism in P pastoris As expected, the f-values obtained for the glucose and glycerol cultivations (Table 2) show that the proteinogenic amino acids are primarily ... Ministry of Science and Technology (CICYT project PPQ2001-1908), and the Academy of Finland (projects 52311 and 202409) The authors thank J M Cregg and C Gancedo for useful comments, and O Cos for ... investigation of amino acid biosynthesis and comparison with flux ratios obtained for growth on glycerol, P pastoris was also grown with glucose as the sole carbon source Materials and methods Strain and media...

Ngày tải lên: 23/03/2014, 12:20

... shading, a second dimer with green and purple shading and a third dimer with blue and yellow shading Red atoms indicate amino acids that dier among B caldolyticus and B subtilis PRibPP synthases ... insert of pJM1 and the original insert of pHO219 (GenBank and EBI Data Bank accession number X83708) revealed three discrepancies Lys289 and Arg294 were found to be glutamic acid and alanine, respectively ... amino acid of B subtilis PRibPP synthase given in parenthesis: Ile43 (Val), Val56 (Cys), Leu70 (Ile), Asn108 (Glu) and Val115 (Phe) Consistent with the surface location of the altered amino acids...

Ngày tải lên: 23/03/2014, 13:20

... N-terminal amino acids, which differ between the human CYP11B1 and CYP11B2 enzyme, and are candidate residues for influencing the enzymatic activity of human aldosterone synthase As the two helices, B and ... aldosterone synthase and the corresponding amino- acid exchanges Nucleotides represented in bold characters indicate mismatched bases in CYP11B2 Codons for the changed amino acids are underlined ... using PROCHECK [33] and PROSA [34] and all the models showed appropriate quality RESULTS Alignment of human, mouse and rat CYP11B1 and CYP11B2 with crystallized cytochromes P450 and human microsomal...

Ngày tải lên: 24/03/2014, 03:21