Tyr51: Key Determinant of the Low Thermostability of the Colwellia psychrerythraea Cold-Shock Protein.


:Cold-shock proteins (Csps) are expressed at lower-than-optimum temperatures, and they function as RNA chaperones; however, no structural studies on psychrophilic Csps have been reported. Here, we aimed to investigate the structure and dynamics of the Csp of psychrophile Colwellia psychrerythraea 34H, ( Cp-Csp). Although Cp-Csp shares sequence homology, common folding patterns, and motifs, including a five β-stranded barrel, with its thermophilic counterparts, its thermostability (37 °C) was markedly lower than those of other Csps. Cp-Csp binds heptathymidine with an affinity of 10-7 M, thereby increasing its thermostability to 50 °C. Nuclear magnetic resonance spectroscopic analysis of the Cp-Csp structure and backbone dynamics revealed a flexible structure with only one salt bridge and 10 residues in the hydrophobic cavity. Notably, Cp-Csp contains Tyr51 instead of the conserved Phe in the hydrophobic core, and its phenolic hydroxyl group projects toward the surface. The Y51F mutation increased the stability of hydrophobic packing and may have allowed for the formation of a K3-E21 salt bridge, thereby increasing its thermostability to 43 °C. Cp-Csp exhibited conformational exchanges in its ribonucleoprotein motifs 1 and 2 (754 and 642 s-1), and heptathymidine binding markedly decreased these motions. Cp-Csp lacks salt bridges and has longer flexible loops and a less compact hydrophobic cavity resulting from Tyr51 compared to mesophilic and thermophilic Csps. These might explain the low thermostability of Cp-Csp. The conformational flexibility of Cp-Csp facilitates its accommodation of nucleic acids at low temperatures in polar oceans and its function as an RNA chaperone for cold adaptation.






Lee Y,Kwak C,Jeong KW,Durai P,Ryu KS,Kim EH,Cheong C,Ahn HC,Kim HJ,Kim Y




Has Abstract


2018-07-03 00:00:00












  • "Molecular switches" on mGluR allosteric ligands that modulate modes of pharmacology.

    abstract::G-protein-coupled receptors (GPCRs) represent the largest class of drug targets, accounting for more than 40% of marketed drugs; however, discovery efforts for many GPCRs have failed to provide viable drug candidates. Historically, drug discovery efforts have focused on developing ligands that act at the orthosteric s...


    pub_type: 杂志文章,评审


    authors: Wood MR,Hopkins CR,Brogan JT,Conn PJ,Lindsley CW

    更新日期:2011-04-05 00:00:00

  • SH2 domain proteins as high-affinity receptor tyrosine kinase substrates.

    abstract::Activation of a growth factor receptor tyrosine kinase (RTK) is accompanied by a rapid autophosphorylation of the receptor on tyrosine residues. Receptor activation has been shown to promote the association of signal-transducing proteins containing SH2 domains (second domain of src homology). These receptor-associated...


    pub_type: 杂志文章


    authors: Sierke SL,Koland JG

    更新日期:1993-09-28 00:00:00

  • High-pressure denaturation of staphylococcal nuclease proline-to-glycine substitution mutants.

    abstract::Our recently reported pressure-jump relaxation kinetics experiments on staphylococcal nuclease folding and unfolding [Vidugiris et al. (1995) Biochemistry 34, 4909] demonstrated that both transitions exhibit positive activation volumes, with that of folding being much larger than that of unfolding. Thus high pressure ...


    pub_type: 杂志文章


    authors: Vidugiris GJ,Truckses DM,Markley JL,Royer CA

    更新日期:1996-03-26 00:00:00

  • Interaction of acetylene and cyanide with the resting state of nitrogenase alpha-96-substituted MoFe proteins.

    abstract::The nitrogenase MoFe protein contains the active site metallocluster called FeMo-cofactor [7Fe-9S-Mo-homocitrate] that exhibits an S = 3/2 EPR signal in the resting state. No interaction with FeMo-cofactor is detected when either substrates or inhibitors are incubated with MoFe protein in the resting state. Rather, th...


    pub_type: 杂志文章


    authors: Benton PM,Mayer SM,Shao J,Hoffman BM,Dean DR,Seefeldt LC

    更新日期:2001-11-20 00:00:00

  • The Cys-Tyr cross-link of cysteine dioxygenase changes the optimal pH of the reaction without a structural change.

    abstract::Cysteine dioxygenase (CDO) is a non-heme monoiron enzyme with an unusual posttranslational modification in the proximity of the ferrous iron active site. This modification, a cysteine to tyrosine thioether bond, cross-links two β-strands of the β-barrel. We have investigated its role in catalysis through a combined cr...


    pub_type: 杂志文章


    authors: Davies CG,Fellner M,Tchesnokov EP,Wilbanks SM,Jameson GN

    更新日期:2014-12-23 00:00:00

  • Spectral studies on the calcium binding properties of bovine brain S-100b protein.

    abstract::The effect of Ca2+ binding on the circular dichroism (CD) and 270-MHz proton nuclear magnetic resonance (NMR) spectra of brain-specific S-100b calcium binding protein has been examined at two pH values, 8.5 and 7.5. At pH 8.5, S-100b protein binds two Ca2+ per monomer with Kd values of 6 x 10(-5) and 2 x 10(-4) M, whe...


    pub_type: 杂志文章


    authors: Mani RS,Shelling JG,Sykes BD,Kay CM

    更新日期:1983-03-29 00:00:00

  • Steroid-protein interactions. Human corticosteroid-binding globulin: characterization of dimer and electrophoretic variants.

    abstract::Human corticosteroid-binding globulin (CBG) forms a dimer that was isolated by gel filtration, has full binding affinity and capacity, and can be dissociated to the monomer. Monomeric CBG consists of two distinct molecular variants, which were detected by polyacrylamide gel electrophoresis in the presence and absence ...


    pub_type: 杂志文章


    authors: Mickelson KE,Harding GB,Forsthoefel M,Westphal U

    更新日期:1982-02-16 00:00:00

  • Aromatic substrate specificity of horseradish peroxidase C studied by a combined fluorescence line narrowing/energy minimization approach: the effect of localized side-chain reorganization.

    abstract::Horseradish peroxidase C binds a wide variety of small H-donor compounds such as benzohydroxamic acid (BHA) and 2-naphthohydroxamic acid (NHA). In this work, we use the Mg(II)-mesoporphyrin prosthetic group derivative as a spectroscopic probe of the active site and of the interaction with the substrates. We report on ...


    pub_type: 杂志文章


    authors: Laberge M,Osvath S,Fidy J

    更新日期:2001-08-07 00:00:00

  • Effect of erythrocyte transbilayer phospholipid distribution on fusion with vesicular stomatitis virus.

    abstract::To identify the specific component(s) in the target membrane involved in fusion of vesicular stomatitis virus (VSV), we examined the interaction of the virus with human erythrocyte membranes with asymmetric and symmetric bilayer distributions of phospholipids. Fusion was monitored spectrofluorometrically by the octade...


    pub_type: 杂志文章


    authors: Herrmann A,Clague MJ,Puri A,Morris SJ,Blumenthal R,Grimaldi S

    更新日期:1990-05-01 00:00:00

  • Crystallographic structures of ribonuclease S variants with nonpolar substitution at position 13: packing and cavities.

    abstract::Seven hydrophobic residues ranging in size from glycine to phenylalanine have been substituted for the wild-type methionine residue at position 13 in a 15-residue truncated version (S15) of S-peptide, the small component of ribonuclease S. Complexes of both S-15 and the seven variants with S-protein yielded isomorphou...


    pub_type: 杂志文章


    authors: Varadarajan R,Richards FM

    更新日期:1992-12-15 00:00:00

  • Exploring the Role of Residue 228 in Substrate and Inhibitor Recognition by VIM Metallo-β-lactamases.

    abstract::β-Lactamase inhibitors (BLIs) restore the efficacy of otherwise obsolete β-lactams. However, commercially available BLIs are not effective against metallo-β-lactamases (MBLs), which continue to be disseminated globally. One group of the most clinically important MBLs is the VIM family. The discovery of VIM-24, a natur...


    pub_type: 杂志文章


    authors: Mojica MF,Mahler SG,Bethel CR,Taracila MA,Kosmopoulou M,Papp-Wallace KM,Llarrull LI,Wilson BM,Marshall SH,Wallace CJ,Villegas MV,Harris ME,Vila AJ,Spencer J,Bonomo RA

    更新日期:2015-05-26 00:00:00

  • Ligand-specific structural changes in the vitamin D receptor in solution.

    abstract::Vitamin D receptor (VDR) is a member of the nuclear hormone receptor superfamily. When bound to a variety of vitamin D analogues, VDR manifests a wide diversity of physiological actions. The molecular mechanism by which different vitamin D analogues cause specific responses is not understood. The published crystallogr...


    pub_type: 杂志文章


    authors: Singarapu KK,Zhu J,Tonelli M,Rao H,Assadi-Porter FM,Westler WM,DeLuca HF,Markley JL

    更新日期:2011-12-27 00:00:00

  • Eukaryotic Catalase-Peroxidase: The Role of the Trp-Tyr-Met Adduct in Protein Stability, Substrate Accessibility, and Catalysis of Hydrogen Peroxide Dismutation.

    abstract::Recently, it was demonstrated that bifunctional catalase-peroxidases (KatGs) are found not only in archaea and bacteria but also in lower eukaryotes. Structural studies and preliminary biochemical data of the secreted KatG from the rice pathogen Magnaporthe grisea (MagKatG2) suggested both similar and novel features w...


    pub_type: 杂志文章


    authors: Gasselhuber B,Carpena X,Graf MM,Pirker KF,Nicolussi A,Sündermann A,Hofbauer S,Zamocky M,Furtmüller PG,Jakopitsch C,Oostenbrink C,Fita I,Obinger C

    更新日期:2015-09-08 00:00:00

  • Bimodal action of alkaline earth cations on Dictyostelium discoideum ribonuclease P activity.

    abstract::The ribonucleoprotein ribonuclease P (RNase P) cleaves all tRNA precursors endonucleolitically to produce the mature 5'-end. Dictyostelium discoideum RNase P displays an absolute requirement for Mg2+. Only the alkaline earth cations Ca2+, Sr2+, and Ba2+, under appropriate conditions can substitute to some extent for M...


    pub_type: 杂志文章


    authors: Tekos A,Stathopoulos C,Drainas D

    更新日期:1998-11-03 00:00:00

  • Endogenous 5-methylcytosine protects neighboring guanines from N7 and O6-methylation and O6-pyridyloxobutylation by the tobacco carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone.

    abstract::All CG dinucleotides along exons 5-8 of the p53 tumor suppressor gene contain endogenous 5-methylcytosine (MeC). These same sites (e.g., codons 157, 158, 245, 248, and 273) are mutational hot spots in smoking-induced lung cancer. Several groups used the UvrABC endonuclease incision assay to demonstrate that methylated...


    pub_type: 杂志文章


    authors: Ziegel R,Shallop A,Upadhyaya P,Jones R,Tretyakova N

    更新日期:2004-01-20 00:00:00

  • DNA intramolecular triplexes containing dT --> dU substitutions: unfolding energetics and ligand binding.

    abstract::We used a combination of optical and calorimetric techniques to investigate the incorporation of deoxythymidine --> deoxyuridine (dT --> dU) substitutions in the duplex and third strand of the parallel intramolecular triplex d(A(7)C(5)T(7)C(5)T(7)) (ATT). UV and differential scanning calorimetry melting experiments sh...


    pub_type: 杂志文章


    authors: Soto AM,Rentzeperis D,Shikiya R,Alonso M,Marky LA

    更新日期:2006-03-07 00:00:00

  • Elimination of the disulfide bridge in the Rieske iron-sulfur protein allows assembly of the [2Fe-2S] cluster into the Rieske protein but damages the ubiquinol oxidation site in the cytochrome bc1 complex.

    abstract::The [2Fe-2S] cluster of the Rieske iron-sulfur protein is held between two loops of the protein that are connected by a disulfide bridge. We have replaced the two cysteines that form the disulfide bridge in the Rieske protein of Saccharomyces cerevisiae with tyrosine and leucine, and tyrosine and valine, to evaluate t...


    pub_type: 杂志文章


    authors: Merbitz-Zahradnik T,Zwicker K,Nett JH,Link TA,Trumpower BL

    更新日期:2003-11-25 00:00:00

  • L-685,458, an aspartyl protease transition state mimic, is a potent inhibitor of amyloid beta-protein precursor gamma-secretase activity.

    abstract::Progressive cerebral amyloid beta-protein (A beta) deposition is believed to play a central role in the pathogenesis of Alzheimer's disease (AD). Elevated levels of A beta(42) peptide formation have been linked to early-onset familial AD-causing gene mutations in the amyloid beta-protein precursor (A beta PP) and the ...


    pub_type: 杂志文章


    authors: Shearman MS,Beher D,Clarke EE,Lewis HD,Harrison T,Hunt P,Nadin A,Smith AL,Stevenson G,Castro JL

    更新日期:2000-08-01 00:00:00

  • cAMP binds with high affinity and specificity to the androgen receptor from murine skeletal muscle.

    abstract::cAMP binding of the androgen receptor (AR) from murine skeletal muscle was studied. Testosterone affinity chromatography yielded androgen receptor with about 4000-fold purification. Determination of the cAMP binding in the affinity eluate, by adsorption of protein-cAMP complexes to cellulose ester filters or removal o...


    pub_type: 杂志文章


    authors: Ofenloch-Hähnle B,Eisele K

    更新日期:1988-11-15 00:00:00

  • Localization of a reactive exofacial sulfhydryl on the glucose carrier of human erythrocytes.

    abstract::Tryptic digestion studies of the human erythrocyte glucose carrier have shown that a reactive and transport-sensitive exofacial sulfhydryl is located in the carboxy-terminal half of the molecule, corresponding to Cys347, Cys421, or Cys429. In the present studies, the erythrocyte glucose carrier labeled on the exofacia...


    pub_type: 杂志文章


    authors: May JM,Buchs A,Carter-Su C

    更新日期:1990-11-13 00:00:00

  • Cyclobutane-type pyrimidine photodimer formation and excision in human skin fibroblasts after irradiation with 313-nm ultraviolet light.

    abstract::The formation and excision of 313-nm light-induced cyclobutane-type pyrimidine photodimers were determined in confluent cultures of human fibroblasts. A new method was developed for the resolution and determination of cytosine-thymine (CT) and thymine-thymine dimers (TT) by using sodium borohydride reduction and high-...


    pub_type: 杂志文章


    authors: Niggli HJ,Cerutti PA

    更新日期:1983-03-15 00:00:00

  • Kinetic studies on the mechanism of pepsin action.

    abstract::The linear noncompetitive inhibition of the pepsin-catalyzed hydrolysis of Ac-Phe-Phe-Gly at pH 2.1 by L-Ac-Phe, L-Ac-Phe-NH2, and L-Ac-Phe-OEt has been claimed to substantiate the ordered release of products specified by the amino-enzyme mechanism for pepsin action. According to this interpretation, the binding of in...


    pub_type: 杂志文章


    authors: Silver MS,Stoddard M

    更新日期:1975-02-11 00:00:00

  • Insights into the mechanism of Pseudomonas dacunhae aspartate beta-decarboxylase from rapid-scanning stopped-flow kinetics.

    abstract::The mechanism of wild-type and R37A mutant Pseudomonas dacunhae aspartate beta-decarboxylase (ABDC) was studied by rapid-scanning stopped-flow spectrophotometry. Mixing wild-type ABDC with 50 mM disodium l-Asp resulted in the formation of a 325 nm absorption peak within the dead time of the stopped-flow instrument, li...


    pub_type: 杂志文章


    authors: Phillips RS,Lima S,Khristoforov R,Sudararaju B

    更新日期:2010-06-22 00:00:00

  • Molecular interactions between human lactotransferrin and the phytohemagglutinin-activated human lymphocyte lactotransferrin receptor lie in two loop-containing regions of the N-terminal domain I of human lactotransferrin.

    abstract::Fluorescein isothiocyanate derivatization of the human lactotransferrin on Lys-264 inhibits the binding of the protein of human PHA-activated lymphocytes [Legrand, D., Mazurier, J., Maes, P., Rochard, E., Montreuil, J., & Spik, G. (1991) Biochem. J. 276, 733-738], indicating that part of the receptor-binding site is l...


    pub_type: 杂志文章


    authors: Legrand D,Mazurier J,Elass A,Rochard E,Vergoten G,Maes P,Montreuil J,Spik G

    更新日期:1992-09-29 00:00:00

  • Reaction of neuronal nitric-oxide synthase with 2,6-dichloroindolphenol and cytochrome c3+: influence of the electron acceptor and binding of Ca2+-activated calmodulin on the kinetic mechanism.

    abstract::Binding of Ca(2+)-activated calmodulin (Ca(2+)-CaM) to neuronal nitric-oxide synthase (nNOS) increases the rate of 2,6-dichloroindolphenol (DCIP) reduction 2-3-fold and that of cytochrome c(3+) 10-20-fold. Parallel initial velocity patterns indicated that both substrates were reduced via two-half reactions in a ping-p...


    pub_type: 杂志文章


    authors: Wolthers KR,Schimerlik MI

    更新日期:2001-04-17 00:00:00

  • Electrostatic guidance of glycosyl cation migration along the reaction coordinate of uracil DNA glycosylase.

    abstract::The DNA repair enzyme uracil DNA glycosylase has been crystallized with a cationic 1-aza-2'-deoxyribose-containing DNA that mimics the ultimate transition state of the reaction in which the water nucleophile attacks the anomeric center of the oxacarbenium ion-uracil anion reaction intermediate. Comparison with substra...


    pub_type: 杂志文章


    authors: Bianchet MA,Seiple LA,Jiang YL,Ichikawa Y,Amzel LM,Stivers JT

    更新日期:2003-11-04 00:00:00

  • Acoustically mounted microcrystals yield high-resolution X-ray structures.

    abstract::We demonstrate a general strategy for determining structures from showers of microcrystals. It uses acoustic droplet ejection to transfer 2.5 nL droplets from the surface of microcrystal slurries, through the air, onto mounting micromesh pins. Individual microcrystals are located by raster-scanning a several-micromete...


    pub_type: 杂志文章


    authors: Soares AS,Engel MA,Stearns R,Datwani S,Olechno J,Ellson R,Skinner JM,Allaire M,Orville AM

    更新日期:2011-05-31 00:00:00

  • Reactivity of essential thiols of myosin. Chemical probes of the activated state.

    abstract::14C-Labeled fluorodinitrobenzene and N-ethylmaleimide have been used as chemical probes of the conformational states of myosin induced by the binding of MgADP and MgATP. The results indicate that in the high-energy conformation, MMgADP-Pi, the essential thiols are protected from modification but their diminished react...


    pub_type: 杂志文章


    authors: Reisler E,Burke M,Harrington WF

    更新日期:1977-11-29 00:00:00

  • Metabolic buffering exerted by macromolecular crowding on DNA-DNA interactions: origin and physiological significance.

    abstract::Crowding, which characterizes the interior of all living cells, has been shown to dramatically affect biochemical processes, leading to stabilization of compact morphologies, enhanced macromolecular associations, and altered reaction rates. Due to the crowding-mediated shift in binding equilibria toward association, c...


    pub_type: 杂志文章


    authors: Goobes R,Kahana N,Cohen O,Minsky A

    更新日期:2003-03-04 00:00:00

  • The CXC motif: a functional mimic of protein disulfide isomerase.

    abstract::Protein disulfide isomerase (PDI) utilizes the active site sequence Cys-Gly-His-Cys (CGHC; E degrees ' = -180 mV) to effect thiol-disulfide interchange during oxidative protein folding. Here, the Cys-Gly-Cys-NH(2) (CGC) peptide is shown to have a disulfide reduction potential (E degrees ' = -167 mV) that is close to t...


    pub_type: 杂志文章


    authors: Woycechowsky KJ,Raines RT

    更新日期:2003-05-13 00:00:00