Role of Asn112 in a Light-Driven Sodium Ion-Pumping Rhodopsin.


:Light-driven outward sodium-pumping rhodopsin (NaR) was recently found in marine bacteria. Krokinobacter eikastus rhodopsin 2 (KR2) actively transports sodium and lithium ions in NaCl and LiCl, respectively, while it pumps protons in KCl. NaR has a conserved NDQ (N112, D116, and Q123 in KR2) motif, and previous studies suggested an important role for N112 in the function of KR2. Here we replaced N112 with 19 different amino acids and studied the molecular properties of the mutants. All mutants exhibited absorption bands from a protonated Schiff base in the λmax range from 508 to 531 nm upon heterologous expression in Escherichia coli, whose ion-pumping activity was measured using pH electrodes. The function of these mutants was classified into three phenotypes: wild-type (WT)-like Na+/H+ compatible pump, exclusive H+ pump, and no pump. Among the 19 mutants, only N112D, -G, -S, and -T showed light-driven Na+ pump activity, N112A, -C, -P, -V, -E, -Q, -I, -L, -M, -F, and -W were exclusively H+ pumps, and N112H, -K, -Y, and -R exhibited no pump activity. The mutants of the no pump function lack a blue-shifted M intermediate, indicating that Schiff base deprotonation is a prerequisite for Na+ and H+ pumps. In contrast, the subsequent red-shifted O intermediate was observed for WT and N112V but absent for N112T and N112A, suggesting that observation of this intermediate depends on kinetics. Although N112D, -G, -S, and -T are able to pump Na+, they also pump H+ in NaCl, where Na+ and H+ pumps compete with each other because of the decreased Na+ uptake efficiency. From these facts, an exclusive Na+ pump in NaCl exists only in WT. We conclude that N112 is one of the functional determinants of NaR.






Abe-Yoshizumi R,Inoue K,Kato HE,Nureki O,Kandori H




Has Abstract


2016-10-18 00:00:00












  • Narrowing substrate specificity in a directly evolved enzyme: the A293D mutant of aspartate aminotransferase.

    abstract::Several mutant Escherichia coli aspartate aminotransferases (eAATases) have been characterized in the attempt to evolve or rationally redesign the substrate specificity of eAATase into that of E. coli tyrosine aminotransferase (eTATase). These include HEX (designed), HEX + A293D (design followed by directed evolution)...


    pub_type: 杂志文章


    authors: Chow MA,McElroy KE,Corbett KD,Berger JM,Kirsch JF

    更新日期:2004-10-12 00:00:00

  • The conserved seven-transmembrane sequence NP(X)2,3Y of the G-protein-coupled receptor superfamily regulates multiple properties of the beta 2-adrenergic receptor.

    abstract::The beta 2-adrenergic receptor (beta 2AR) is a member of a large superfamily of seven transmembrane domain, G-protein-coupled receptors. Within the putative seventh transmembrane domain of the beta 2AR is a sequence of amino acids, NPLIY, which is conserved with minor variations in all members of the superfamily. Prev...


    pub_type: 杂志文章


    authors: Barak LS,Ménard L,Ferguson SS,Colapietro AM,Caron MG

    更新日期:1995-11-28 00:00:00

  • Trihydroxynaphthalene reductase from Magnaporthe grisea: realization of an active center inhibitor and elucidation of the kinetic mechanism.

    abstract::Active trihydroxynaphthalene reductase (3HNR) is essential for the biosynthesis of fungal melanin by Magnaporthe grisea and is a focus of inhibitor design studies directed toward control of blast disease in rice. Tricyclazole, a preventative fungicide against rice blast, has been previously characterized as inhibiting...


    pub_type: 杂志文章


    authors: Thompson JE,Basarab GS,Andersson A,Lindqvist Y,Jordan DB

    更新日期:1997-02-18 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

  • Low molecular weight protein tyrosine phosphatase and caveolin-1: interaction and isoenzyme-dependent regulation.

    abstract::Low molecular weight protein tyrosine phosphatases (LMW-PTPs) are small enzymes that are ubiquitous in many organisms. They are important in biological processes such as cell proliferation, adhesion, migration, and invasiveness. LMW-PTP is expressed in mammalian cells as two isoforms (IF1 and IF2) originating through ...


    pub_type: 杂志文章


    authors: Caselli A,Taddei ML,Bini C,Paoli P,Camici G,Manao G,Cirri P,Ramponi G

    更新日期:2007-05-29 00:00:00

  • Role of mitochondrial and cytoplasmic serine hydroxymethyltransferase isozymes in de novo purine synthesis in Saccharomyces cerevisiae.

    abstract::One-carbon units are essential to a variety of anabolic processes which yield necessary cellular components including purines, pyrimidines, amino acids, and lipids. Serine hydroxymethyltransferase (SHMT) is the major provider of one-carbon units in the cell. The other product of this reaction is glycine. Both of these...


    pub_type: 杂志文章


    authors: Kastanos EK,Woldman YY,Appling DR

    更新日期:1997-12-02 00:00:00

  • Turnover and orientation of the major neural retina cell surface protein protected from tryptic cleavage by calcium.

    abstract::Calcium protects a limited number of embryonic chick neural retina cell surface proteins from tryptic cleavage. One glycoprotein of Mr approximately 1.3 X 10(5) and pI approximately 4.8 (gp130) is present in intact retinas and, in the presence of Ca2+, is both resistant to tryptic cleavage and poorly iodinated. When c...


    pub_type: 杂志文章


    authors: Cook JH,Pratt RS,Lilien J

    更新日期:1984-02-28 00:00:00

  • Involvement of Phenylalanine 297 in the Construction of the Substrate Pocket of Human Aminopeptidase B.

    abstract::Aminopeptidase B (APB, EC preferentially hydrolyzes the N-terminal basic amino acids of synthetic and peptide substrates and requires a physiological concentration of NaCl for optimal activity. In this study, we used site-directed mutagenesis and molecular modeling to search for an amino acid residue that is...


    pub_type: 杂志文章


    authors: Ohnishi A,Watanabe J,Ogawa Y,Goto Y,Hattori A,Tsujimoto M

    更新日期:2015-10-06 00:00:00

  • Thrombospondin-1 and angiotensin II inhibit soluble guanylyl cyclase through an increase in intracellular calcium concentration.

    abstract::Nitric oxide (NO) regulates cardiovascular hemostasis by binding to soluble guanylyl cyclase (sGC), leading to cGMP production, reduced cytosolic calcium concentration ([Ca(2+)](i)), and vasorelaxation. Thrombospondin-1 (TSP-1), a secreted matricellular protein, was recently discovered to inhibit NO signaling and sGC ...


    pub_type: 杂志文章


    authors: Ramanathan S,Mazzalupo S,Boitano S,Montfort WR

    更新日期:2011-09-13 00:00:00

  • Certain metal ions are inhibitors of cytochrome b6f complex 'Rieske' iron-sulfur protein domain movements.

    abstract::Many current models of the Q cycle for the cytochrome (cyt) b6f and the cyt bc1 complexes incorporate 'Rieske' iron-sulfur protein (ISP) domain movements to gate electron transfer and to ensure high yields of proton shuttling. It was previously proposed that copper ions, which bind at a site distant from the quinol ox...


    pub_type: 杂志文章


    authors: Roberts AG,Bowman MK,Kramer DM

    更新日期:2002-03-26 00:00:00

  • An additional copy of the homologous region (hr1) sequence in the Autographa californica multinucleocapsid polyhedrosis virus genome promotes hyperexpression of foreign genes.

    abstract::The Autographa californica multinucleocapsid nuclear polyhedrosis virus genome contains nine homologous region (hr1, hr1a, hr2, hr2a, hr3, hr4a, hr4b, hr4c, and hr5) sequences that are thought to be involved in viral replication and activation of transcription. Our results show that the 750 bp hr1 sequence is capable ...


    pub_type: 杂志文章


    authors: Venkaiah B,Viswanathan P,Habib S,Hasnain SE

    更新日期:2004-06-29 00:00:00

  • Structural determinants of protein dynamics: analysis of 15N NMR relaxation measurements for main-chain and side-chain nuclei of hen egg white lysozyme.

    abstract::15N-labeled hen lysozyme has been studied by 2D and 3D NMR in order to characterize its dynamic behavior. The resonances of all main-chain amide nitrogen atoms were assigned, as were resonances of nitrogen atoms in 28 side chains. Relaxation measurements for the main-chain and arginine and tryptophan side-chain 15N nu...


    pub_type: 杂志文章


    authors: Buck M,Boyd J,Redfield C,MacKenzie DA,Jeenes DJ,Archer DB,Dobson CM

    更新日期:1995-03-28 00:00:00

  • One-Enzyme Reverse Transcription qPCR Using Taq DNA Polymerase.

    abstract::Taq DNA polymerase, one of the first thermostable DNA polymerases to be discovered, has been typecast as a DNA-dependent DNA polymerase commonly employed for PCR. However, Taq polymerase belongs to the same DNA polymerase superfamily as the Molony murine leukemia virus reverse transcriptase and has in the past been sh...


    pub_type: 杂志文章


    authors: Bhadra S,Maranhao AC,Paik I,Ellington AD

    更新日期:2020-12-15 00:00:00

  • Circadian genomics reveal a role for post-transcriptional regulation in mammals.

    abstract::To maintain daily cycles, the circadian clock must tightly regulate the rhythms of thousands of mRNAs and proteins with the correct period, phase, and amplitude to ultimately drive the wide range of rhythmic biological processes. Recent genomic approaches have revolutionized our view of circadian gene expression and h...


    pub_type: 杂志文章,评审


    authors: Kojima S,Green CB

    更新日期:2015-01-20 00:00:00

  • Insight into the catalysis of hydrolysis of four newly synthesized substrates by papain: a proton inventory study.

    abstract::We synthesized the following four new peptide substrates, Suc-Phe-Leu-pNA, Suc-Phe-Leu-NMec, Suc-Phe-Leu-ONPh, and Pht-Phe-Leu-pNA, and we applied the proton inventory method to their hydrolysis by papain. Useful relationships between the rate constants of the catalytic reaction have been established and contributed t...


    pub_type: 杂志文章


    authors: Theodorou LG,Lymperopoulos K,Bieth JG,Papamichael EM

    更新日期:2001-04-03 00:00:00

  • Secondary structure and interaction of phage D108 Ner repressor with a 61-base-pair operator: evidence for altered protein and DNA structures in the complex.

    abstract::Ner repressors of the transposable phages Mu and D108 play a central role in regulating the expression of the early (transposase) operon and in ensuring that phage growth proceeds along a lytic pathway. The latter function is analogous to that performed by the Cro protein of phage lambda. Unlike lambda Cro, however, t...


    pub_type: 杂志文章


    authors: Benevides JM,Kukolj G,Autexier C,Aubrey KL,DuBow MS,Thomas GJ Jr

    更新日期:1994-09-06 00:00:00

  • Neurofibrillar tangle surrogates: histone H1 binding to patterned phosphotyrosine peptide nanotubes.

    abstract::Living cells contain a range of densely phosphorylated surfaces, including phospholipid membranes, ribonucleoproteins, and nucleic acid polymers. Hyperphosphorylated surfaces also accumulate in neurodegenerative diseases as neurofibrillar tangles. We have synthesized and structurally characterized a precisely patterne...


    pub_type: 杂志文章


    authors: Li S,Sidorov AN,Mehta AK,Bisignano AJ,Das D,Childers WS,Schuler E,Jiang Z,Orlando TM,Berland K,Lynn DG

    更新日期:2014-07-08 00:00:00

  • Identification of c-di-GMP derivatives resistant to an EAL domain phosphodiesterase.

    abstract::The bacterial second messenger signaling molecule bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) controls important biological processes such as biofilm formation, virulence response, and motility. This second messenger is sensed by macromolecular targets inside the cell, both protein and RNA, which ind...


    pub_type: 杂志文章


    authors: Shanahan CA,Gaffney BL,Jones RA,Strobel SA

    更新日期:2013-01-15 00:00:00

  • High-yield cleavage of tryptophanyl peptide bonds by o-iodosobenzoic acid.

    abstract::A new procedure to cleave tryptophanyl peptide bonds in high yield is reported. The method involves treatment of the S-alkylated protein with o-iodosobenzoic acid. The procedure is highly selective for tryptophan and does not modify tyrosine or histidine, but may convert methionine to its sulfoxide derivative. The yie...


    pub_type: 杂志文章


    authors: Mahoney WC,Hermodson MA

    更新日期:1979-08-21 00:00:00

  • Kinetics of intracomplex electron transfer and of reduction of the components of covalent and noncovalent complexes of cytochrome c and cytochrome c peroxidase by free flavin semiquinones.

    abstract::The kinetics of reduction of free flavin semiquinones of the individual components of 1:1 covalent and electrostatic complexes of yeast ferric and ferryl cytochrome c peroxidase and ferric horse cytochrome c have been studied. Covalent cross-linking between the peroxidase and cytochrome c at low ionic strength results...


    pub_type: 杂志文章


    authors: Hazzard JT,Moench SJ,Erman JE,Satterlee JD,Tollin G

    更新日期:1988-03-22 00:00:00

  • DNA sequence-specific recognition by the Saccharomyces cerevisiae "TATA" binding protein: promoter-dependent differences in the thermodynamics and kinetics of binding.

    abstract::The equilibrium binding and association kinetics of the Saccharomyces cerevisiae TATA Binding Protein (TBP) to the E4 and Major Late promoters of adenovirus (TATATATA and TATAAAAG, respectively), have been directly compared by quantitative DNase I titration and quench-flow "footprinting". The equilibrium binding of TB...


    pub_type: 杂志文章


    authors: Petri V,Hsieh M,Jamison E,Brenowitz M

    更新日期:1998-11-10 00:00:00

  • Tyrosine kinase-mediated serine phosphorylation of adenylyl cyclase.

    abstract::Receptor tyrosine kinase (RTK) activation is associated with modulation of heptahelical receptor-stimulated adenylyl cyclase responses. The mechanisms underlying the RTK-mediated enhancement of adenylyl cyclase function remain unclear. In the present studies, we show that the tyrosine kinase-dependent enhancement of a...


    pub_type: 杂志文章


    authors: Tan CM,Kelvin DJ,Litchfield DW,Ferguson SS,Feldman RD

    更新日期:2001-02-13 00:00:00

  • Identification by proton nuclear magnetic resonance of the histidines in cytochrome b5 modified by diethyl pyrocarbonate.

    abstract::Diethyl pyrocarbonate (DEP) is an electrophilic reagent that is used to modify reversibly the histidine residues of proteins. Unfortunately, the lability of the acylated histidine adduct usually does not permit the isolation and identification of the modified histidine. By use of 500-MHz proton NMR spectroscopy, it ha...


    pub_type: 杂志文章


    authors: Altman J,Lipka JJ,Kuntz I,Waskell L

    更新日期:1989-09-19 00:00:00

  • Dimers to doughnuts: redox-sensitive oligomerization of 2-cysteine peroxiredoxins.

    abstract::2-Cys peroxiredoxins (Prxs) are a large and diverse family of peroxidases which, in addition to their antioxidant functions, regulate cell signaling pathways, apoptosis, and differentiation. These enzymes are obligate homodimers (alpha(2)), utilizing a unique intermolecular redox-active disulfide center for the reduct...


    pub_type: 杂志文章


    authors: Wood ZA,Poole LB,Hantgan RR,Karplus PA

    更新日期:2002-04-30 00:00:00

  • The Influenza M2 Ectodomain Regulates the Conformational Equilibria of the Transmembrane Proton Channel: Insights from Solid-State Nuclear Magnetic Resonance.

    abstract::The influenza M2 protein is the target of the amantadine family of antiviral drugs, and its transmembrane (TM) domain structure and dynamics have been extensively studied. However, little is known about the structure of the highly conserved N-terminal ectodomain, which contains epitopes targeted by influenza vaccines....


    pub_type: 杂志文章


    authors: Kwon B,Hong M

    更新日期:2016-09-27 00:00:00

  • Stoichiometry of LTbetaR binding to LIGHT.

    abstract::LTbetaR is a member of the TNF receptor family of proteins. It binds to two different cell surface ligands, LIGHT, a homotypic trimer, and LTalpha1beta2, a heterotypic trimer. We have measured the affinities of the dimeric IgG fusion protein, LTbetaRIgG, and monomeric LTbetaR protein binding to both LIGHT and LTalpha1...


    pub_type: 杂志文章


    authors: Eldredge J,Berkowitz S,Corin AF,Day ES,Hayes D,Meier W,Strauch K,Zafari M,Tadi M,Farrington GK

    更新日期:2006-08-22 00:00:00

  • Plant Translation Initiation Complex eIFiso4F Directs Pokeweed Antiviral Protein to Selectively Depurinate Uncapped Tobacco Etch Virus RNA.

    abstract::Pokeweed antiviral protein (PAP) is a ribosome inactivating protein (RIP) that depurinates the sarcin/ricin loop (SRL) of rRNA, inhibiting protein synthesis. PAP depurinates viral RNA, and in doing so, lowers the infectivity of many plant viruses. The mechanism by which PAP accesses uncapped viral RNA is not known, im...


    pub_type: 杂志文章


    authors: Domashevskiy AV,Williams S,Kluge C,Cheng SY

    更新日期:2017-11-14 00:00:00

  • Differential coupling of smooth and skeletal muscle pyruvate kinase to creatine kinase.

    abstract::The interaction of pyruvate kinase from skeletal (SKPK) and smooth (SMPK) muscle with MM-creatine kinase (MMCK) and BB-creatine kinase (BBCK) was assessed using temporal absorbance changes, variations in absorbance at different wavelengths, concentration dependence, association in an electric field, and PK kinetic act...


    pub_type: 杂志文章


    authors: Sears PR,Dillon PF

    更新日期:1999-11-09 00:00:00

  • Molecular exchange at the lipid-rhodopsin interface: spin-label electron spin resonance studies of rhodopsin-dimyristoylphosphatidylcholine recombinants.

    abstract::The photoreceptor protein rhodopsin has been reconstituted with a single phospholipid species, dimyristoylphosphatidylcholine, at a range of different lipid/protein ratios, and the exchange rate at the lipid-protein interface has been determined from the electron spin resonance spectra of spin-labeled phosphatidylchol...


    pub_type: 杂志文章


    authors: Ryba NJ,Horváth LI,Watts A,Marsh D

    更新日期:1987-06-02 00:00:00

  • Redox properties of electron-transfer flavoprotein ubiquinone oxidoreductase as determined by EPR-spectroelectrochemistry.

    abstract::We have determined the formal potential values for each electron transfer to electron transfer flavoprotein-ubiquinone oxidoreductase (ETF-QO), in order to further characterize the thermodynamics of electron transport from various acyl-CoA thioesters to the mitochondrial ubiquinone pool. ETF-QO contains one [4Fe-4S]2+...


    pub_type: 杂志文章


    authors: Paulsen KE,Orville AM,Frerman FE,Lipscomb JD,Stankovich MT

    更新日期:1992-12-01 00:00:00