Acquisition of an insertion peptide for efficient aminoacylation by a halophile tRNA synthetase.


:Enzymes of halophilic organisms contain unusual peptide motifs that are absent from their mesophilic counterparts. The functions of these halophile-specific peptides are largely unknown. Here we have identified an unusual peptide that is unique to several halophile archaeal cysteinyl-tRNA synthetases (CysRS), which catalyze attachment of cysteine to tRNA(Cys) to generate the essential cysteinyl-tRNA(Cys) required for protein synthesis. This peptide is located near the active site in the catalytic domain and is highly enriched with acidic residues. In the CysRS of the extreme halophile Halobacterium species NRC-1, deletion of the peptide reduces the catalytic efficiency of aminoacylation by a factor of 100 that largely results from a defect in kcat, rather than the Km for tRNA(Cys). In contrast, maintaining the peptide length but substituting acidic residues in the peptide with neutral or basic residues has no major deleterious effect, suggesting that the acidity of the peptide is not important for the kcat of tRNA aminoacylation. Analysis of general protein structure under physiological high salt concentrations, by circular dichroism and by fluorescence titration of tRNA binding, indicates little change due to deletion of the peptide. However, the presence of the peptide confers tolerance to lower salt levels, and fluorescence analysis in 30% sucrose reveals instability of the enzyme without the peptide. We suggest that the stability associated with the peptide can be used to promote proper enzyme conformation transitions in various stages of tRNA aminoacylation that are associated with catalysis. The acquisition of the peptide by the halophilic CysRS suggests an enzyme adaptation to high salinity.






Evilia C,Hou YM




Has Abstract


2006-06-06 00:00:00












  • Purification and characterization of CTP synthetase, the product of the URA7 gene in Saccharomyces cerevisiae.

    abstract::In the yeast Saccharomyces cerevisiae, CTP synthetase [EC; UTP:ammonia ligase (ADP-forming)] is the product of the URA7 gene. CTP synthetase was purified 503-fold to apparent homogeneity from cells bearing the URA7 gene on a multicopy plasmid that directed a 10-fold overproduction of the enzyme. The purificati...


    pub_type: 杂志文章


    authors: Yang WL,McDonough VM,Ozier-Kalogeropoulos O,Adeline MT,Flocco MT,Carman GM

    更新日期:1994-09-06 00:00:00

  • Glutathione induces helical formation in the carboxy terminus of human glutathione transferase A1-1.

    abstract::The structure and dynamic properties of the C-terminal region of the human class alpha glutathione transferase A1-1 have been investigated with high-resolution NMR methods. On the basis of crystallographic and fluorescence measurements, this 13-residue segment of the enzyme is presumed to be disordered in the unligand...


    pub_type: 杂志文章


    authors: Zhan Y,Rule GS

    更新日期:2004-06-15 00:00:00

  • Thermodynamic characterization of ligand-induced conformational changes in UDP-N-acetylglucosamine enolpyruvyl transferase.

    abstract::The binding of UDP-N-acetylglucosamine (UDPNAG) to the enzyme UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) was studied in the absence and presence of the antibiotic fosfomycin by isothermal titration calorimetry. Fosfomycin binds covalently to MurA in the presence of UDPNAG and also in its absence as demonst...


    pub_type: 杂志文章


    authors: Samland AK,Jelesarov I,Kuhn R,Amrhein N,Macheroux P

    更新日期:2001-08-21 00:00:00

  • Model for general acid-base catalysis by the hammerhead ribozyme: pH-activity relationships of G8 and G12 variants at the putative active site.

    abstract::We have used nucleobase substitution and kinetic analysis to test the hypothesis that hammerhead catalysis occurs by a general acid-base mechanism, in which nucleobases are directly involved in deprotonation of the attacking 2'-hydroxyl group and protonation of the 5'-oxygen that serves as the leaving group in the cle...


    pub_type: 杂志文章


    authors: Han J,Burke JM

    更新日期:2005-05-31 00:00:00

  • Allosteric activation of protein phosphatase 2C by D-chiro-inositol-galactosamine, a putative mediator mimetic of insulin action.

    abstract::Insulin-stimulated glucose disposal in skeletal muscle proceeds predominantly through a nonoxidative pathway with glycogen synthase as a rate-limiting enzyme, yet the mechanisms for insulin activation of glycogen synthase are not understood despite years of investigation. Isolation of putative insulin second messenger...


    pub_type: 杂志文章


    authors: Brautigan DL,Brown M,Grindrod S,Chinigo G,Kruszewski A,Lukasik SM,Bushweller JH,Horal M,Keller S,Tamura S,Heimark DB,Price J,Larner AN,Larner J

    更新日期:2005-08-23 00:00:00

  • Human Cytosolic and Mitochondrial Serine Hydroxymethyltransferase Isoforms in Comparison: Full Kinetic Characterization and Substrate Inhibition Properties.

    abstract::Serine hydroxymethyltransferase (SHMT) catalyzes the reversible conversion of l-serine and tetrahydrofolate into glycine and 5,10-methylenetetrahydrofolate. This enzyme, which plays a pivotal role in one-carbon metabolism, is involved in cancer metabolic reprogramming and is a recognized target of chemotherapy interve...


    pub_type: 杂志文章


    authors: Tramonti A,Nardella C,di Salvo ML,Barile A,Cutruzzolà F,Contestabile R

    更新日期:2018-12-26 00:00:00

  • Triplex formation of chemically modified homopyrimidine oligonucleotides: thermodynamic and kinetic studies.

    abstract::We have investigated effects of chemical modifications of a third strand on the thermodynamic and kinetic properties of the triplex formation between a 23-bp duplex and each of four kinds of 15-mer chemically modified third strands using isothermal titration calorimetry and interaction analysis system. The chemical mo...


    pub_type: 杂志文章


    authors: Torigoe H,Shimizume R,Sarai A,Shindo H

    更新日期:1999-11-02 00:00:00

  • The metastasis-associated extracellular matrix protein osteopontin forms transient structure in ligand interaction sites.

    abstract::Osteopontin (OPN) is an acidic hydrophilic glycophosphoprotein that was first identified as a major sialoprotein in bones. It functions as a cell attachment protein displaying a RGD cell adhesion sequence and as a cytokine that signals through integrin and CD44 cell adhesion molecules. OPN is also implicated in human ...


    pub_type: 杂志文章


    authors: Platzer G,Schedlbauer A,Chemelli A,Ozdowy P,Coudevylle N,Auer R,Kontaxis G,Hartl M,Miles AJ,Wallace BA,Glatter O,Bister K,Konrat R

    更新日期:2011-07-12 00:00:00

  • Thermodynamic parameters of the binding of retinol to binding proteins and to membranes.

    abstract::Retinol (vitamin A alcohol) is a hydrophobic compound and distributes in vivo mainly between binding proteins and cellular membranes. To better clarify the nature of the interactions of retinol with these phases which have a high affinity for it, the thermodynamic parameters of these interactions were studied. The tem...


    pub_type: 杂志文章


    authors: Noy N,Xu ZJ

    更新日期:1990-04-24 00:00:00

  • Determination of disulfide structure in agouti-related protein (AGRP) by stepwise reduction and alkylation.

    abstract::The agouti-related protein gene (Agrp) plays an important role in body weight regulation. The mature human protein is a single polypeptide chain of 112 amino acid residues, consisting of an N-terminal acidic region and a unique C-terminal cysteine-rich domain. The disulfide structure of recombinant human AGRP was dete...


    pub_type: 杂志文章


    authors: Bures EJ,Hui JO,Young Y,Chow DT,Katta V,Rohde MF,Zeni L,Rosenfeld RD,Stark KL,Haniu M

    更新日期:1998-09-01 00:00:00

  • Synthesis and properties of azidonitrophenyl pyrophosphate, a photoaffinity probe of the nucleotide binding sites of mitochondrial F1-ATPase.

    abstract::4-Azido-2-nitrophenyl pyrophosphate (azido-PPi) labeled with 32P in the alpha position was prepared and used to photolabel beef heart mitochondrial F1. Azido-PPi was hydrolyzed by yeast inorganic pyrophosphatase, but not by mitochondrial F1-ATPase. Incubation of F1 with [alpha-32P]azido-PPi in the dark under condition...


    pub_type: 杂志文章


    authors: Michel L,Garin J,Issartel JP,Vignais PV

    更新日期:1989-12-26 00:00:00

  • Pre-steady-state studies of phosphite dehydrogenase demonstrate that hydride transfer is fully rate limiting.

    abstract::Phosphite dehydrogenase (PTDH) is a unique NAD-dependent enzyme that catalyzes the oxidation of inorganic phosphite to phosphate. The enzyme has great potential for cofactor regeneration, and mechanistic studies have provided some insight into the residues that are important for catalysis. In this investigation, pre-s...


    pub_type: 杂志文章


    authors: Fogle EJ,van der Donk WA

    更新日期:2007-11-13 00:00:00

  • Environmental influences on the in vivo level of intramolecular triplex DNA in Escherichia coli.

    abstract::We describe an assay for detecting intramolecular triple-stranded DNA in living cells. The assay involves quantitative analysis of the differential photobinding of 4,5',8-trimethylpsoralen to 5'TA and 5'AT dinucleotides in a region of plasmid DNA containing the triplex-forming sequence (GA)7TA(GA)7. Psoralen photobind...


    pub_type: 杂志文章


    authors: Ussery DW,Sinden RR

    更新日期:1993-06-22 00:00:00

  • Two distinct mechanisms for regulation of nonmuscle myosin assembly via the heavy chain: phosphorylation for MIIB and mts 1 binding for MIIA.

    abstract::In search of the regulation mechanisms for isoform specific myosin assembly, we have used the COOH-terminal fragments of nonmuscle myosin isoforms MIIA and MIIB (MIIA(F46) and MIIB(alpha)(F47)) as a model system. Phosphorylation by protein kinase C (PK C) or casein kinase II (CK II) within or near the nonhelical tail-...


    pub_type: 杂志文章


    authors: Murakami N,Kotula L,Hwang YW

    更新日期:2000-09-19 00:00:00

  • The dimerization stability of the HLH-LZ transcription protein family is modulated by the leucine zippers: a CD and NMR study of TFEB and c-Myc.

    abstract::In the HLH-LZ protein family, the helix-loop-helix DNA-binding dimerization domain is followed in the sequence by a leucine zipper motif. The precise function of this second dimerization domain is still unclear, since the HLH motif of a subset of this family has been shown to be necessary and sufficient for dimerizati...


    pub_type: 杂志文章


    authors: Muhle-Goll C,Gibson T,Schuck P,Schubert D,Nalis D,Nilges M,Pastore A

    更新日期:1994-09-20 00:00:00

  • Circular dichroism studies on the interaction of tryptophan synthase with pyridoxal 5'-phosphate.

    abstract::The interaction between the beta 2 subunit of tryptophan synthase and the coenzyme pyridoxal 5'-phosphate (PLP) is characterized by induced circular dichroism (CD) in the near-UV (260-285 nm) and in the visible region (320-480 nm, extrinsic Cotton effect). Because of its high mean residue ellipticity ([theta] = 56 deg...


    pub_type: 杂志文章


    authors: Balk H,Merkl I,Bartholmes P

    更新日期:1981-10-27 00:00:00

  • Structure of the potassium form of CGCGAATTCGCG: DNA deformation by electrostatic collapse around inorganic cations.

    abstract::The potassium form of d(CGCGAATTCGCG) solved by X-ray diffraction to 1.75 A resolution indicates that monovalent cations penetrate the primary and secondary layers of the "spine of hydration". Both the sodium [Shui, X., McFail-Isom, L., Hu, G. G., and Williams, L. D. (1998) Biochemistry 37, 8341-8355] and the potassiu...


    pub_type: 杂志文章


    authors: Shui X,Sines CC,McFail-Isom L,VanDerveer D,Williams LD

    更新日期:1998-12-01 00:00:00

  • Binding of a photoaffinity analogue of glutathione to MRP1 (ABCC1) within two cytoplasmic regions (L0 and L1) as well as transmembrane domains 10-11 and 16-17.

    abstract::MRP1 (or ABCC1) is an ABC membrane protein that transports a wide range of natural products as well as glutathione (GSH)-, glucuronate-, and sulfate-conjugated metabolites. In addition, free GSH is required for MRP1 to transport several chemotherapeutic drugs. However, the mechanisms regulating the influence of GSH on...


    pub_type: 杂志文章


    authors: Karwatsky J,Daoud R,Cai J,Gros P,Georges E

    更新日期:2003-03-25 00:00:00

  • An expanded two-state model accounts for homotropic cooperativity in biosynthetic threonine deaminase from Escherichia coli.

    abstract::The linkage between substrate and regulatory effector binding to separate sites on allosteric enzymes results in shifts in their sigmoidal kinetics to regulate metabolism. Control of branched chain amino acid biosynthesis in Escherichia coli occurs in part through shifts in the sigmoidal dependence of alpha-ketobutyra...


    pub_type: 杂志文章


    authors: Eisenstein E,Yu HD,Fisher KE,Iacuzio DA,Ducote KR,Schwarz FP

    更新日期:1995-07-25 00:00:00

  • Olfactory transduction: cross-talk between second-messenger systems.

    abstract::Chemosensory cilia of olfactory receptor neurons contain an adenylate cyclase which is stimulated by high concentrations of odorants. Cyclic AMP produced by this enzyme has been proposed to act as second messenger in olfactory transduction. Here we report that olfactory cilia contain calmodulin and that calmodulin pot...


    pub_type: 杂志文章


    authors: Anholt RR,Rivers AM

    更新日期:1990-05-01 00:00:00

  • Binding of cytochrome P450 monooxygenase and lipoxygenase pathway products by heart fatty acid-binding protein.

    abstract::Arachidonic acid metabolism by lipoxygenases and cytochrome P450 monooxygenases produces regioisomeric hydroperoxyeicosatetraenoic acids (HPETEs), hydroxyeicosatetraenoic acids (HETEs), epoxyeicosatrienoic acids (EETs), and dihydroxyeicosatrienoic acids (DHETs), which serve as components of cell signaling cascades. In...


    pub_type: 杂志文章


    authors: Widstrom RL,Norris AW,Spector AA

    更新日期:2001-01-30 00:00:00

  • Physical dissection of the structural elements responsible for regulatory properties and intersubunit interactions of protein kinase CK2 beta-subunit.

    abstract::The noncatalytic beta-subunit of protein kinase CK2 has been shown to display various and in some respects antagonistic effects on the catalytic alpha-subunit [Boldyreff et al. (1993) Biochemistry 32, 12672-12677; Meggio et al. (1994) Biochemistry 33, 4336-4342]. We have now examined the ability of peptides encompassi...


    pub_type: 杂志文章


    authors: Marin O,Meggio F,Sarno S,Pinna LA

    更新日期:1997-06-10 00:00:00

  • Evidence for a nucleotide binding site on the isolated beta subunit from Escherichia coli F1-ATPase. Interaction between nucleotide and aurovertin D binding sites.

    abstract::The nucleotide binding capacity and affinity of the isolated beta subunit from Escherichia coli F1-ATPase have been studied with radiolabeled ADP and ATP by an equilibrium dialysis technique. Each mole of beta subunit in the presence of EDTA bound 1 mol of ADP or ATP with Kd values of 25 microM and 50-100 microM, resp...


    pub_type: 杂志文章


    authors: Issartel JP,Vignais PV

    更新日期:1984-12-18 00:00:00

  • Evolution of the antibiotic resistance protein, FosA, is linked to a catalytically promiscuous progenitor.

    abstract::The fosfomycin (1) resistance proteins FosA and FosX in pathogenic microorganisms are related to a catalytically promiscuous progenitor encoded in a phn operon in Mesorhizobium loti. The mlr3345 gene product (FosX(Ml)) from M. loti has a very low epoxide hydrolase activity and even lower glutathione transferase activi...


    pub_type: 杂志文章


    authors: Brown DW,Schaab MR,Birmingham WR,Armstrong RN

    更新日期:2009-03-10 00:00:00

  • Catalytic function and local proton structure at the type 2 copper of nitrite reductase: the correlation of enzymatic pH dependence, conserved residues, and proton hyperfine structure.

    abstract::Electron nuclear double resonance (ENDOR) of protons at Type 2 and Type 1 cupric active sites correlates with the enzymatic pH dependence, the mutation of nearby conserved, nonligating residues, and electron transfer in heterologously expressed Rhodobacter sphaeroides nitrite reductase. Wild-type enzyme showed a pH 6 ...


    pub_type: 杂志文章


    authors: Zhao Y,Lukoyanov DA,Toropov YV,Wu K,Shapleigh JP,Scholes CP

    更新日期:2002-06-11 00:00:00

  • Rho transcription factor: symmetry and binding of bicyclomycin.

    abstract::The antibiotic bicyclomycin inhibits rho-dependent termination processes by interfering with RNA translocation by preventing RNA binding at the translocation site or by uncoupling the translocation process from ATP hydrolysis. Previous studies have shown that bicyclomycin binds near the ATP hydrolysis pocket on rho. T...


    pub_type: 杂志文章


    authors: Vincent F,Openshaw M,Trautwein M,Gaskell SJ,Kohn H,Widger WR

    更新日期:2000-08-08 00:00:00

  • Photoreactions of the Histidine Kinase Rhodopsin Ot-HKR from the Marine Picoalga Ostreococcus tauri.

    abstract::The tiny picoalga, Ostreococcus tauri, originating from the Thau Lagoon is a member of the marine phytoplankton. Because of its highly reduced genome and small cell size, while retaining the fundamental requirements of a eukaryotic photosynthetic cell, it became a popular model organism for studying photosynthesis or ...


    pub_type: 杂志文章


    authors: Luck M,Velázquez Escobar F,Glass K,Sabotke MI,Hagedorn R,Corellou F,Siebert F,Hildebrandt P,Hegemann P

    更新日期:2019-04-09 00:00:00

  • Characterization of the tertiary structure of soluble CD4 bound to glycosylated full-length HIVgp120 by chemical modification of arginine residues and mass spectrometric analysis.

    abstract::The initial step of infection of blood cells with the human immunodeficiency virus, HIV, is the formation of a complex of the viral envelope protein gp120 and its human receptor CD4. We have examined structural features of recombinant soluble CD4 (sCD4) by chemical modification of arginine residues with hydroxyphenylg...


    pub_type: 杂志文章


    authors: Hager-Braun C,Tomer KB

    更新日期:2002-02-12 00:00:00

  • Communications between the high-affinity cyclic nucleotide binding sites in E. coli cyclic AMP receptor protein: effect of single site mutations.

    abstract::The binding of adenosine 3',5'-cyclic monophosphate (cAMP) and its nonfunctional analogue, guanosine 3',5'-cyclic monophosphate (cGMP), to the adenosine 3',5'-cyclic monophosphate receptor protein (CRP) from Escherichia coli was investigated by means of fluorescence and isothermal titration calorimetry (ITC) at pH 7.8...


    pub_type: 杂志文章


    authors: Lin SH,Lee JC

    更新日期:2002-10-01 00:00:00

  • Control of the chemical step by leucine-31 of pancreatic phospholipase A2.

    abstract::A well-defined region of pancreatic and other secreted phospholipase A2 (PLA2), which we call the i-face, makes a molecular contact with the interface to facilitate and control the events and processivity of the interfacial catalytic turnover cycles. The structural features of the i-face and its allosteric relationshi...


    pub_type: 杂志文章


    authors: Yu BZ,Janssen MJ,Verheij HM,Jain MK

    更新日期:2000-05-16 00:00:00