Completely buried, non-ion-paired glutamic acid contributes favorably to the conformational stability of pyrrolidone carboxyl peptidases from hyperthermophiles.


:Pyrrolidone carboxyl peptidases (PCPs) from hyperthermophiles have a structurally conserved and completely buried Glu192 in the hydrophobic core; in contrast, the corresponding residue in the mesophile protein is a hydrophobic residue, Ile. Does the buried ionizable residue contribute to stabilization or destabilization of hyperthermophile PCPs? To elucidate the role of the buried glutamic acid in stabilizing PCP from hyperthermophiles, we constructed five Glu192 mutants of PCP-0SH (C142S/C188S, Cys-free double mutant of PCP) from Pyrococcus furiosus and examined their thermal and pH-induced unfolding and crystal structures and compared them with those of PCP-0SH. The stabilities of apolar (E192A/I/V) and polar (E192D/Q) mutants were less than PCP-0SH at acidic pH values. In the alkaline region, the mutant proteins, except for E192D, were more stable than PCP-0SH. The thermal stability data and theoretical calculations indicated an apparent pKa value > or = 7.3 for Glu192. Present results confirmed that the protonated Glu192 in PCP-0SH forms strong hydrogen bonds with the carbonyl oxygen and peptide nitrogen of Pro168. New intermolecular hydrogen bonds in the E --> A/D mutants were formed by a water molecule introduced into the cavity created around position 192, whereas the hydrogen bonds disappeared in the E --> I/V mutants. Structure-based empirical stability of mutant proteins was in good agreement with the experimental results. The results indicated that (1) completely buried Glu192 contributes to the stabilization of PCP-0SH because of the formation of strong intramolecular hydrogen bonds and (2) the hydrogen bonds by the nonionized and buried Glu can contribute more than the burial of hydrophobic groups to the conformational stability of proteins.






Kaushik JK,Iimura S,Ogasahara K,Yamagata Y,Segawa S,Yutani K




Has Abstract


2006-06-13 00:00:00












  • Catalase HPII from Escherichia coli exhibits enhanced resistance to denaturation.

    abstract::Catalase HPII from Escherichia coli is a homotetramer of 753 residue subunits. The multimer displays a number of unusual structural features, including interwoven subunits and a covalent bond between Tyr415 and His392, that would contribute to its rigidity and stability. As the temperature of a solution of HPII in 50 ...


    pub_type: 杂志文章


    authors: Switala J,O'Neil JO,Loewen PC

    更新日期:1999-03-30 00:00:00

  • Solution structure of the Dickerson DNA dodecamer containing a single ribonucleotide.

    abstract::Ribonucleotides are frequently incorporated into DNA during replication. They are recognized and processed by several cellular enzymes, and their continued presence in the yeast nuclear genome results in replicative stress and genome instability. Thus, it is important to understand the effects of isolated ribonucleoti...


    pub_type: 杂志文章


    authors: DeRose EF,Perera L,Murray MS,Kunkel TA,London RE

    更新日期:2012-03-27 00:00:00

  • Fate of the (2Fe-2S)(2+) cluster of Escherichia coli biotin synthase during reaction: a Mössbauer characterization.

    abstract::Biotin synthase, the enzyme which catalyzes the last step of the biosynthesis of biotin, contains only (2Fe-2S)(2+) clusters when isolated under aerobic conditions. Previous results showed that reduction by dithionite or photoreduced deazaflavin converts the (2Fe-2S)(2+) to (4Fe-4S)(2+,+). However, until now, no detai...


    pub_type: 杂志文章


    authors: Tse Sum Bui B,Benda R,Schünemann V,Florentin D,Trautwein AX,Marquet A

    更新日期:2003-07-29 00:00:00

  • Characterization of Natural Colibactin-Nucleobase Adducts by Tandem Mass Spectrometry and Isotopic Labeling. Support for DNA Alkylation by Cyclopropane Ring Opening.

    abstract::Colibactins are genotoxic secondary metabolites whose biosynthesis is encoded in the clb gene cluster harbored by certain strains of gut commensal Escherichia coli. Using synthetic colibactin analogues, we previously provided evidence that colibactins alkylate DNA by addition of a nucleotide to an electrophilic cyclop...


    pub_type: 杂志文章


    authors: Xue M,Shine E,Wang W,Crawford JM,Herzon SB

    更新日期:2018-11-13 00:00:00

  • Role of the covalent flavin linkage in monomeric sarcosine oxidase.

    abstract::Monomeric sarcosine oxidase (MSOX) is a prototypical member of a recently recognized family of amine-oxidizing enzymes that all contain covalently bound flavin. Mutation of the covalent flavin attachment site in MSOX produces a catalytically inactive apoprotein (apoCys315Ala) that forms an unstable complex with FAD (K...


    pub_type: 杂志文章


    authors: Hassan-Abdallah A,Zhao G,Jorns MS

    更新日期:2006-08-08 00:00:00

  • Mapping the interfacial binding surface of human secretory group IIa phospholipase A2.

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    pub_type: 杂志文章


    authors: Snitko Y,Koduri RS,Han SK,Othman R,Baker SF,Molini BJ,Wilton DC,Gelb MH,Cho W

    更新日期:1997-11-25 00:00:00

  • Gross structural changes in isolated liver cell plasma membranes upon binding of insulin.

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    pub_type: 杂志文章


    authors: Luly P,Shinitzky M

    更新日期:1979-02-06 00:00:00

  • Estimation of the distance change between cysteine-457 and the nucleotide binding site when sodium pump changes conformation from E1 to E2 by fluorescence energy transfer measurements.

    abstract::The first indication of the size of a conformational change implicated in ion transport by sodium pump has been obtained by measuring the change in efficiency of fluorescence energy transfer between two specific locations on the alpha-subunit. The donor (5'-(iodoacetamido)fluorescein) attaches covalently to cysteine-4...


    pub_type: 杂志文章


    authors: Lin SH,Faller LD

    更新日期:1996-06-25 00:00:00

  • Kinetics of the subtransition of asymmetrically substituted phosphatidylcholines.

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    pub_type: 杂志文章


    authors: Tümmler B,Herrmann U,Maass G,Eibl H

    更新日期:1984-08-28 00:00:00

  • A-tract polarity dominate the curvature in flanking sequences.

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    pub_type: 杂志文章


    authors: Merling A,Sagaydakova N,Haran TE

    更新日期:2003-05-06 00:00:00

  • In Situ Photoregulation of Carbonic Anhydrase Activity Using Azobenzenesulfonamides.

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    pub_type: 杂志文章


    authors: Aggarwal K,Banik M,Medellin B,Que EL

    更新日期:2019-01-08 00:00:00

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    abstract::The transporter associated with antigen processing (TAP) is a prototype of an asymmetric ATP-binding cassette (ABC) transporter, which uses ATP binding and hydrolysis to translocate peptides from the cytosol to the lumen of the endoplasmic reticulum (ER). Here, we review molecular details of peptide binding and ATP bi...


    pub_type: 杂志文章,评审


    authors: Hinz A,Tampé R

    更新日期:2012-06-26 00:00:00

  • Inactivation of C30A trimethylamine dehydrogenase by N-cyclopropyl-alpha-methylbenzylamine, 1-phenylcyclopropylamine, and phenylhydrazine.

    abstract::Trimethylamine dehydrogenase (TMADH) from the bacterium Methylophilus methylotrophus (sp. W(3)A(1)) and its C30A mutant were inactivated with three known inactivators of monoamine oxidase, namely, phenylhydrazine, N-cyclopropyl-alpha-methylbenzylamine, and 1-phenylcyclopropylamine. All three compounds irreversibly ina...


    pub_type: 杂志文章


    authors: Mitchell DJ,Nikolic D,Jang MH,van Breemen RB,Hille R,Silverman RB

    更新日期:2001-07-24 00:00:00

  • An engineered disulfide bond in dihydrofolate reductase.

    abstract::Substitution of cysteine for proline-39 in Escherichia coli dihydrofolate reductase by oligonucleotide-directed mutagenesis positions the new cysteine adjacent to already existing cysteine-85. When the mutant protein is expressed in the E. coli cytosol, the cysteine sulfur atoms are found, by X-ray crystallographic an...


    pub_type: 杂志文章


    authors: Villafranca JE,Howell EE,Oatley SJ,Xuong NH,Kraut J

    更新日期:1987-04-21 00:00:00

  • An Efficient Viologen-Based Electron Donor to Nitrogenase.

    abstract::Nitrogenase catalyzes the reduction of N2 to NH3, supporting all biological nitrogen fixation. Electron donors to this enzyme are ferredoxin or flavodoxin (in vivo) and sodium dithionite (in vitro). Features of these electron donors put a limit on spectrophotometric studies and electrocatalytic applications of nitroge...


    pub_type: 杂志文章


    authors: Badalyan A,Yang ZY,Hu B,Luo J,Hu M,Liu TL,Seefeldt LC

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    authors: Schalk IJ,Kyslik P,Prome D,van Dorsselaer A,Poole K,Abdallah MA,Pattus F

    更新日期:1999-07-20 00:00:00

  • Regulation of the expression of the β-lactam antibiotic-resistance determinants in methicillin-resistant Staphylococcus aureus (MRSA).

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    pub_type: 杂志文章


    authors: Blázquez B,Llarrull LI,Luque-Ortega JR,Alfonso C,Boggess B,Mobashery S

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    pub_type: 杂志文章


    authors: Kim DW,Huang T,Schirch D,Schirch V

    更新日期:1996-12-10 00:00:00

  • HlyC, the internal protein acyltransferase that activates hemolysin toxin: role of conserved histidine, serine, and cysteine residues in enzymatic activity as probed by chemical modification and site-directed mutagenesis.

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    pub_type: 杂志文章


    authors: Trent MS,Worsham LM,Ernst-Fonberg ML

    更新日期:1999-03-16 00:00:00

  • Structure-function relationships in human glutathione-dependent formaldehyde dehydrogenase. Role of Glu-67 and Arg-368 in the catalytic mechanism.

    abstract::The active-site zinc in human glutathione-dependent formaldehyde dehydrogenase (FDH) undergoes coenzyme-induced displacement and transient coordination to a highly conserved glutamate residue (Glu-67) during the catalytic cycle. The role of this transient coordination of the active-site zinc to Glu-67 in the FDH catal...


    pub_type: 杂志文章


    authors: Sanghani PC,Davis WI,Zhai L,Robinson H

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  • Solid-State Nuclear Magnetic Resonance Structural Study of the Retinal-Binding Pocket in Sodium Ion Pump Rhodopsin.

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    pub_type: 杂志文章


    authors: Shigeta A,Ito S,Inoue K,Okitsu T,Wada A,Kandori H,Kawamura I

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    pub_type: 杂志文章


    authors: Nicoletti FP,Comandini A,Bonamore A,Boechi L,Boubeta FM,Feis A,Smulevich G,Boffi A

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  • Cadmium-113 NMR studies of the DNA binding domain of the mammalian glucocorticoid receptor.

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    pub_type: 杂志文章


    authors: Pan T,Freedman LP,Coleman JE

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  • Interaction of C225SR1 mutant subunit of ribonucleotide reductase with R2 and nucleoside diphosphates: tales of a suicidal enzyme.

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    pub_type: 杂志文章


    authors: Mao SS,Holler TP,Bollinger JM Jr,Yu GX,Johnston MI,Stubbe J

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    pub_type: 杂志文章


    authors: Olson MO,Kirstein MN,Wallace MO

    更新日期:1990-06-19 00:00:00

  • Conformational Heterogeneity in a Fully Complementary DNA Three-Way Junction with a GC-Rich Branchpoint.

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    pub_type: 杂志文章


    authors: Toulmin A,Baltierra-Jasso LE,Morten MJ,Sabir T,McGlynn P,Schröder GF,Smith BO,Magennis SW

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  • Characterization of myotoxin a from the venom of prairie rattlesnake (Crotalus viridis viridis).

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    pub_type: 杂志文章


    authors: Cameron DL,Tu AT

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    pub_type: 杂志文章


    authors: Yamasaki R,Bacon BE,Nasholds W,Schneider H,Griffiss JM

    更新日期:1991-10-29 00:00:00

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    pub_type: 杂志文章


    authors: Iliescu A,Gravel M,Horth C,Apuzzo S,Gros P

    更新日期:2011-03-29 00:00:00

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    pub_type: 杂志文章


    authors: Nadal RC,Davies P,Brown DR,Viles JH

    更新日期:2009-09-29 00:00:00