Critical nature of a specific uridine O2-carbonyl for cleavage by the hammerhead ribozyme.

Abstract:

:Three modified hammerhead ribozyme/substrate complexes have been prepared in which individual uridine O2-carbonyls have been eliminated. The modified complexes were chemically synthesized with the substitution of a single 2-pyridone (2P) base analogue for residues U4, U7, and U16.1. Steady-state kinetic analyses indicate that the cleavage efficiencies for the U7 and U16.1 complexes were not significantly reduced relative to the native complex as measured by kcat/KM. The cleavage efficiency for the 2P4 complex, with the analogue present within the uridine loop, was reduced by greater than 2 orders of magnitude. This significant reduction in catalytic efficiency was due primarily to a decrease in kcat. The pH vs cleavage rate profile suggests that the O2-carbonyl of the U4 residue of the hammerhead complex is critical for transition state stabilization and efficient cleavage activity. The results of a Mg2+ rescue assay do not implicate the O2-carbonyl of U4 in an interaction with a divalent metal ion. In addition, the results of a ribozyme folding assay suggest that the presence of the 2P4 within the uridine loop does not alter the folding pathway (relative to the native sequence) both in the absence and in the presence of Mg2+. The O2-carbonyl of U4 appears oriented toward the interior of the catalytic pocket where it may be involved in a critical hydrogen bonding interaction necessary for transition state stabilization.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Bevers S,Ha SB,McLaughlin LW

doi

10.1021/bi982977a

subject

Has Abstract

pub_date

1999-06-15 00:00:00

pages

7710-8

issue

24

eissn

0006-2960

issn

1520-4995

pii

bi982977a

journal_volume

38

pub_type

杂志文章
  • Homodinuclear (Pt,Pt) and heterodinuclear (Ru,Pt) metal compounds as DNA-protein cross-linking agents: potential suicide DNA lesions.

    abstract::Homodinuclear (Pt,Pt) and heterodinuclear (Ru,Pt) metal compounds having the generalized formula M(a)NH2(CH)4NH2M(b) are shown to form specific DNA lesions which can efficiently cross-link proteins to DNA. In this study, the homodinuclear case is represented by M(a) = M(b) = [cis-Pt(Cl2)-(NH3)] and the heterodinuclear...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00095a007

    authors: Van Houten B,Illenye S,Qu Y,Farrell N

    更新日期:1993-11-09 00:00:00

  • Abortive initiation and long ribonucleic acid synthesis.

    abstract::In vitro transcription assays have been used to study the rate of ribonucleic acid (RNA) synthesis from the Escherichia coli lactose promotor mutant lacL8UV5 contained on a 203-bp (base pair) restriction fragment. The half-life of long (63-base) RNA production from heparin-resistant RNA polymerase-promotor complexes w...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00511a003

    authors: Munson LM,Reznikoff WS

    更新日期:1981-04-14 00:00:00

  • Substrate-induced inactivation of a crippled beta-glucosidase mutant: identification of the labeled amino acid and mutagenic analysis of its role.

    abstract::The beta-glucosidase from Agrobacterium sp. catalyzes the hydrolysis of beta-glucosides via a covalent alpha-D-glucopyranosyl-enzyme intermediate involving Glu358. Hydrolysis of 2,4-dinitrophenyl beta-D-glucopyranoside by the low activity Glu358Asp mutant of Agrobacterium beta-glucosidase is accompanied by time-depend...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00044a033

    authors: Gebler JC,Trimbur DE,Warren AJ,Aebersold R,Namchuk M,Withers SG

    更新日期:1995-11-07 00:00:00

  • Photoreactivities and thermal properties of psoralen cross-links.

    abstract::We have studied the photoreaction of 8-methoxypsoralen (8-MOP), 4,5',8-trimethylpsoralen (TMP), and 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT) with a pair of 18-base-long oligonucleotides in which a 14-base region is complementary. Only one 5'TpA site, favored for both monoadduct and cross-link formation with p...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00409a012

    authors: Yeung AT,Jones BK,Chu CT

    更新日期:1988-05-03 00:00:00

  • Organization of the human protein S genes.

    abstract::Human genomic clones that span the entire protein S expressed gene (PS alpha) and the 3' two-thirds of the protein S pseudogene (PS beta) have been isolated and characterized. The PS alpha gene is greater than 80 kilobases in length and contains 14 introns and 15 exons, as well as 6 repetitive "Alu" sequences. Exons I...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00486a010

    authors: Schmidel DK,Tatro AV,Phelps LG,Tomczak JA,Long GL

    更新日期:1990-08-28 00:00:00

  • Novel transcriptional activities of vitamin E: inhibition of cholesterol biosynthesis.

    abstract::Vitamin E is a dietary lipid that is essential for vertebrate health and fertility. The biological activity of vitamin E is thought to reflect its ability to quench oxygen- and carbon-based free radicals and thus to protect the organism from oxidative damage. However, recent reports suggest that vitamin E may also dis...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi701432q

    authors: Valastyan S,Thakur V,Johnson A,Kumar K,Manor D

    更新日期:2008-01-15 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...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi026993q

    authors: Woycechowsky KJ,Raines RT

    更新日期:2003-05-13 00:00:00

  • Interaction of the antitumor compound cryptophycin-52 with tubulin.

    abstract::Cryptophycin-52 (LY355703) is currently undergoing clinical evaluation for cancer chemotherapy. It is a potent suppressor of microtubule dynamics in vitro, and low picomolar concentrations appear to inhibit cancer cell proliferation at mitosis by stabilizing spindle microtubules. In the present study, using [(3)H]cryp...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi0010827

    authors: Panda D,Ananthnarayan V,Larson G,Shih C,Jordan MA,Wilson L

    更新日期:2000-11-21 00:00:00

  • Modification of the conformational equilibria in the sodium and potassium dependent adenosinetriphosphatase with glutaraldehyde.

    abstract::Glutaraldehyde treatment of electroplax membrane preparations of Na,K-ATPase leads to irreversible changes in the enzymic behavior of the protein, which are not due to modification of the active site. When the glutaraldehyde treatment is carried out in a medium containing K+ and without Na+, the "K+-modified enzyme" s...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00288a016

    authors: Chipman DM,Lev A

    更新日期:1983-09-13 00:00:00

  • Effects of sterol structure and exogenous lipids on the transbilayer distribution of sterols in the membrane of Mycoplasma capricolum.

    abstract::Stopped-flow kinetic measurements of the association of filipin with sterols in intact cells and isolated membranes of Mycoplasma capricolum were used to study the effects of varying the phospholipid in the membrane. The phospholipid composition and content of the membrane were varied by growing cells in an albumin-co...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00511a019

    authors: Clejan S,Bittman R,Rottem S

    更新日期:1981-04-14 00:00:00

  • Asymmetric Roles of Two Histidine Residues in Streptococcus pyogenes Cas9 Catalytic Domains upon Chemical Rescue.

    abstract::CRISPR-Cas9 technology has been at the forefront of the field of biology. The Streptococcus pyogenes (SpyCas9) protein forms a complex with guide RNA and can recognize and cleave double-stranded DNA through hybridization based on 20 base pairings. SpyCas9 has two nuclease domains, HNH and RuvC, each of which cuts each...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.0c00766

    authors: Furuhata Y,Kato Y

    更新日期:2021-01-26 00:00:00

  • Revisiting the Val/Ile Mutation in Mammalian and Bacterial Nitric Oxide Synthases: A Spectroscopic and Kinetic Study.

    abstract::Nitric oxide is produced in mammals by the nitric oxide synthase (NOS) isoforms at a catalytic site comprising a heme associated with a biopterin cofactor. Through genome sequencing, proteins that are highly homologous to the oxygenase domain of NOSs have been identified, in particular in bacteria. The active site is ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.6b01018

    authors: Weisslocker-Schaetzel M,Lembrouk M,Santolini J,Dorlet P

    更新日期:2017-02-07 00:00:00

  • Protein arginine deiminase 4: evidence for a reverse protonation mechanism.

    abstract::The presumed role of an overactive protein arginine deiminase 4 (PAD4) in the pathophysiology of rheumatoid arthritis (RA) suggests that PAD4 inhibitors could be used to treat an underlying cause of RA, potentially offering a mechanism to stop further disease progression. Thus, the development of such inhibitors is of...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi700095s

    authors: Knuckley B,Bhatia M,Thompson PR

    更新日期:2007-06-05 00:00:00

  • Relaxing the nicotinamide cofactor specificity of phosphite dehydrogenase by rational design.

    abstract::Homology modeling was used to identify two particular residues, Glu175 and Ala176, in Pseudomonas stutzeri phosphite dehydrogenase (PTDH) as the principal determinants of nicotinamide cofactor (NAD(+) and NADP(+)) specificity. Replacement of these two residues by site-directed mutagenesis with Ala175 and Arg176 both s...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi035018b

    authors: Woodyer R,van der Donk WA,Zhao H

    更新日期:2003-10-14 00:00:00

  • Design, Synthesis, and Potency of Pyruvate Dehydrogenase Complex E1 Inhibitors against Cyanobacteria.

    abstract::Safe and effective algaecides are needed to control agriculturally and environmentally significant algal species. Four series (6, 10, 17, and 21) of 29 novel 4-aminopyrimidine derivatives were rationally designed and synthesized. A part of 10, 17, and 21 displayed potent inhibition of Escherichia coli pyruvate dehydro...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.7b00636

    authors: Zhou Y,Feng J,He H,Hou L,Jiang W,Xie D,Feng L,Cai M,Peng H

    更新日期:2017-12-12 00:00:00

  • Structural Control of Nonnative Ligand Binding in Engineered Mutants of Phosphoenolpyruvate Carboxykinase.

    abstract::Protein engineering to alter recognition underlying ligand binding and activity has enormous potential. Here, ligand binding for Escherichia coli phosphoenolpyruvate carboxykinase (PEPCK), which converts oxaloacetate into CO2 and phosphoenolpyruvate as the first committed step in gluconeogenesis, was engineered to acc...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.8b00963

    authors: Tang HYH,Shin DS,Hura GL,Yang Y,Hu X,Lightstone FC,McGee MD,Padgett HS,Yannone SM,Tainer JA

    更新日期:2018-12-04 00:00:00

  • The subcellular distribution of adenylate and guanylate cyclases in murine lymphoid cells.

    abstract::Membrane vesicles can be prepared from murine lymphoid cells by nitrogen cavitation and fractionated by sedimentation through nonlinear sucrose density gradients. Two subpopulations of membrane vesicles, PMI and PMII, can be distinguished on the basis of sedimentation rate. The subcellular distribution of adenylate an...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00652a026

    authors: Watson J,Nilsen-Hamilton M,Hamilton RT

    更新日期:1976-04-06 00:00:00

  • Conformational and aggregational properties of the gene 9 minor coat protein of bacteriophage M13 in membrane-mimicking systems.

    abstract::The membrane-bound state of the gene 9 minor coat protein of bacteriophage M13 was studied in various membrane-mimicking systems, including organic solvents, detergent micelles, and phospholipid bilayers. For this purpose we determined the conformational and aggregational properties of the chemically synthesized prote...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi981149e

    authors: Houbiers MC,Spruijt RB,Wolfs CJ,Hemminga MA

    更新日期:1999-01-19 00:00:00

  • Structural, Functional, and Dynamical Responses of a Protein in a Restricted Environment Imposed by Macromolecular Crowding.

    abstract::The intercellular environment is known to be very different from the environment where most of the elementary biological processes are studied in the laboratory. As a result, there was a considerable effort on cell mimicking either by confinement or by introducing macromolecular crowding. In the present study, dextran...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.8b00599

    authors: Das N,Sen P

    更新日期:2018-10-23 00:00:00

  • Variable-temperature spectroelectrochemical study of horseradish peroxidase.

    abstract::The reduction potentials of the compound II/ferric and compound I/compound II couples have been studied, using potassium hexachloroiridate as a mediator titrant, by thin-layer spectroelectrochemistry. Compound I, which is 2 equiv more oxidized than the ferric (i.e., resting) form of the enzyme, was reversibly formed v...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00009a017

    authors: Farhangrazi ZS,Fossett ME,Powers LS,Ellis WR Jr

    更新日期:1995-03-07 00:00:00

  • Mechanistic studies on full length and the catalytic domain of the tandem SH2 domain-containing protein tyrosine phosphatase: analysis of phosphoenzyme levels and Vmax stimulatory effects of glycerol and of a phosphotyrosyl peptide ligand.

    abstract::SHP-1, a protein tyrosine phosphatase containing two tandem SH2 domains, is autoinhibited at rest by its N-terminal SH2 (N-SH2) domain. Relief from autoinhibition and a subsequent 10-60-fold increase in V(max) have been observed upon N-SH2 domain engagement by a specific phosphotyrosyl ligand or upon deletion of the S...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9611873

    authors: Wang J,Walsh CT

    更新日期:1997-03-11 00:00:00

  • Activity and crystal structure of Arabidopsis thaliana UDP-N-acetylglucosamine acyltransferase.

    abstract::The UDP-N-acetylglucosamine (UDP-GlcNAc) acyltransferase, encoded by lpxA, catalyzes the first step of lipid A biosynthesis in Gram-negative bacteria, the (R)-3-hydroxyacyl-ACP-dependent acylation of the 3-OH group of UDP-GlcNAc. Recently, we demonstrated that the Arabidopsis thaliana orthologs of six enzymes of the b...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi3002242

    authors: Joo SH,Chung HS,Raetz CR,Garrett TA

    更新日期:2012-05-29 00:00:00

  • Amino acid structures of multiple forms of amyloid-related serum protein SAA from a single individual.

    abstract::Multiple forms of the acute-phase serum protein SAA were isolated from the lipoprotein fraction of plasma from a single individual. These protein forms were purified by size-exclusion, ion-exchange, and reverse-phase high-pressure liquid chromatography, and then the tryptic peptides were subjected to amino acid sequen...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00405a044

    authors: Dwulet FE,Wallace DK,Benson MD

    更新日期:1988-03-08 00:00:00

  • Influence of cofactor pyridoxal 5'-phosphate on reversible high-pressure denaturation of isolated beta 2 dimer of tryptophan synthase bienzyme complex from Escherichia coli.

    abstract::High hydrostatic pressure has been shown to cause reversible dissociation of the isolated apo beta 2 dimer of tryptophan synthase from Escherichia coli into enzymatically inactive monomers [Seifert, T., Bartholmes, P., & Jaenicke, R. (1982) Biophys. Chem. 15, 1-8]. Addition of the coenzyme pyridoxal 5'-phosphate affec...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00323a016

    authors: Seifert T,Bartholmes P,Jaenicke R

    更新日期:1985-01-15 00:00:00

  • Effect of cholesterol on the membrane interaction of Modelin-5 isoforms.

    abstract::Modelin-5 isoforms were used to gain an insight into the effects of amidation on antimicrobial selectivity. When tested against Escherichia coli, amidation increased toxicity 10-fold (MIC = 31.25 μM) while showing limited increased hemolytic activity (2% lysis). Our results show that both the amidated and non-amidated...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi201267v

    authors: Dennison SR,Phoenix DA

    更新日期:2011-12-20 00:00:00

  • The selectivity for K+ versus Na+ in DNA quadruplexes is dominated by relative free energies of hydration: a thermodynamic analysis by 1H NMR.

    abstract::We have studied the competition between Na+ and K+ for coordination by G quartets using the oligonucleotide d(G3T4G3) as a model system. d(G3T4G3) forms a dimeric foldback structure containing three G quartets in the presence of either NaCl or KCl. Proton chemical shifts, which are particular to the species of coordin...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9620565

    authors: Hud NV,Smith FW,Anet FA,Feigon J

    更新日期:1996-12-03 00:00:00

  • Interactions of poly(N epsilon, N epsilon, N epsilon-trimethyllysine) and poly(N delta, N delta, N delta-trimethylornithine) with polynucleotides: salt dissociation and thermal denaturation.

    abstract::The interaction of poly(N epsilon, N epsilon, N epsilon-trimethyl-L-lysine) ([Lys(Me3)]n) and poly(N delta, N delta, N delta-trimethyl-L-ornithine) ([Orn(Me3)]n) with polynucleotides was studied by thermal denaturation, viscosity, and dissociation by salt. The methylated polymers decrease the viscosity of DNA in propo...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00555a020

    authors: Granados EN,Bello J

    更新日期:1980-07-08 00:00:00

  • A conserved threonine within Escherichia coli leucyl-tRNA synthetase prevents hydrolytic editing of leucyl-tRNALeu.

    abstract::Aminoacyl-tRNA synthetases ensure the fidelity of protein synthesis by accurately selecting and activating cognate amino acids for aminoacylation of the correct tRNA. Some tRNA synthetases have evolved an editing active site that is separate from the amino acid activation site providing two steps or "sieves" for amino...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi002915w

    authors: Mursinna RS,Lincecum TL Jr,Martinis SA

    更新日期:2001-05-08 00:00:00

  • The application of 0.1 M quadrol to the microsequence of proteins and the sequence of tryptic peptides.

    abstract::In an effort to extend automated Edman degradation to nanomole quantities of protein, the method of sequenator analysis described by Edman and Begg (Edman, P., and Begg, G. (1967), Eur. J. Biochem. 1, 80) was modified to permit long degradations in the absence of carrier proteins. By using an aqueous 0.1 M Quadrol pro...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00684a036

    authors: Brauer AW,Margolies MN,Haber E

    更新日期:1975-07-01 00:00:00

  • Simple purification and functional reconstitution of octopus photoreceptor Gq, which couples rhodopsin to phospholipase C.

    abstract::In invertebrate photoreceptors, illuminated rhodopsin activates multiple G proteins, which are assumed to initiate multiple phototransduction cascades. In this paper, we focused on one of the phototransduction cascades, which utilizes rhodopsin, a Gq-like G protein, and phospholipase C (PLC). A Gq-like G protein from ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi961360v

    authors: Kikkawa S,Tominaga K,Nakagawa M,Iwasa T,Tsuda M

    更新日期:1996-12-10 00:00:00