Synchronization of the three reaction centers within carbamoyl phosphate synthetase.

Abstract:

:Carbamoyl phosphate synthetase from E. coli catalyzes the synthesis of carbamoyl phosphate through a series of four reactions occurring at three active sites connected by a molecular tunnel of 100 A. To understand the mechanism for coordination and synchronization among the active sites, the pre-steady-state time courses for the formation of phosphate, ADP, glutamate, and carbamoyl phosphate were determined. When bicarbonate and ATP were rapidly mixed with CPS, a stoichiometric burst of acid-labile phosphate and ADP was observed with a formation rate constant of 1100 min(-)(1). The burst phase was followed by a linear steady-state phase with a rate constant of 12 min(-)(1). When glutamine or ammonia was added to the initial reaction mixture, the magnitude and the rate of formation of the burst phase for either phosphate or ADP were unchanged, but the rate constant for the linear steady-state phase increased to an average value of 78 min(-)(1). These results demonstrate that the initial phosphorylation of bicarbonate is independent of the binding or hydrolysis of glutamine. The pre-steady-state time course for the hydrolysis of glutamine in the absence of ATP exhibited a burst of glutamate formation with a rate constant of 4 min(-)(1) when the reaction was quenched with base. In the presence of ATP and bicarbonate, the rate constant for the formation of the burst of glutamate was 1100 min(-)(1). The hydrolysis of ATP thus enhanced the hydrolysis of glutamine by a factor of 275, but there was no effect by glutamine on the initial phosphorylation of bicarbonate. The pre-steady-state time course for the formation of carbamoyl phosphate was linear with an overall rate constant of 72 min(-)(1). The absence of an initial burst of carbamoyl phosphate formation eliminates product release as a rate-determining step for CPS. Overall, these results have been interpreted to be consistent with a mechanism whereby the phosphorylation of bicarbonate serves as the initial trigger for the rest of the reaction cascade. The formation of the carboxy phosphate intermediate within the large subunit must induce a conformational change to the active site of the small subunit that enhances the hydrolysis of glutamine. Thus, ammonia is not released into the molecular tunnel until the activated bicarbonate is ready to form carbamate. The rate-limiting step for the steady-state assembly of carbamoyl phosphate is either the formation, migration, or phosphorylation of the carbamate intermediate.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Miles BW,Raushel FM

doi

10.1021/bi992772h

subject

Has Abstract

pub_date

2000-05-02 00:00:00

pages

5051-6

issue

17

eissn

0006-2960

issn

1520-4995

pii

bi992772h

journal_volume

39

pub_type

杂志文章
  • 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

  • Long-range interaction networks in the function and fidelity of poliovirus RNA-dependent RNA polymerase studied by nuclear magnetic resonance.

    abstract::The fidelity of the poliovirus RNA-dependent RNA polymerase (3D(pol)) plays a direct role in the genomic evolution and pathogenesis of the virus. A single site mutation (Gly64Ser) that is remote from the catalytic center results in a higher fidelity polymerase. NMR studies with [methyl-(13)C]methionine-labeled protein...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi100833r

    authors: Yang X,Welch JL,Arnold JJ,Boehr DD

    更新日期:2010-11-02 00:00:00

  • Clamp-loader-helicase interaction in Bacillus. Leucine 381 is critical for pentamerization and helicase binding of the Bacillus tau protein.

    abstract::Recently, we revealed the architecture of the clamp-loader-helicase (tau-DnaB) complex in Bacillus by atomic force microscopy imaging and constructed a structural model, whereby a pentameric clamp-loader interacts with the hexameric helicase. Crucial to this model is the assumption that the clamp-loader forms a pentam...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi034955g

    authors: Haroniti A,Till R,Smith MC,Soultanas P

    更新日期:2003-09-23 00:00:00

  • Effect of insulin on ATP-citrate lyase phosphorylation: regulation of peptide A and peptide B phosphorylations.

    abstract::Insulin decreases multifunctional protein kinase (MFPK) activity in rat adipose tissue [Ramakrishna, S., & Benjamin, W. B. (1988) J. Biol. Chem. 263, 12677-12681]. Insulin also decreases the phosphorylation of peptide B but increases the phosphorylation of peptide A of ATP-citrate lyase (ATP-CL). The mechanism for thi...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00428a067

    authors: Ramakrishna S,Murthy KS,Benjamin WB

    更新日期:1989-01-24 00:00:00

  • Labeling of two different regions of the nucleotide binding site of the uncoupling protein from brown adipose tissue mitochondria with two ATP analogs.

    abstract::The nucleotide binding site of the uncoupling protein (UCP) from brown adipose tissue was mapped by photoaffinity labeling with 2-azidoadenosine 5'-triphosphate (2-azido-ATP) and by affinity labeling with 3'-O-(5-fluoro-2,4-dinitrophenyl)adenosine 5'-triphosphate (FDNP-ATP). Both analogs bind with high affinity and sp...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00158a017

    authors: Mayinger P,Klingenberg M

    更新日期:1992-11-03 00:00:00

  • Comparison of the Forward and Reverse Photocycle Dynamics of Two Highly Similar Canonical Red/Green Cyanobacteriochromes Reveals Unexpected Differences.

    abstract::Cyanobacteriochromes (CBCRs) are cyanobacterial photoreceptors that exhibit photochromism between two states: a thermally stable dark-adapted state and a metastable light-adapted state with bound linear tetrapyrrole (bilin) chromophores possessing 15Z and 15E configurations, respectively. The photodynamics of canonica...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.0c00796

    authors: Kirpich JS,Chang CW,Franse J,Yu Q,Escobar FV,Jenkins AJ,Martin SS,Narikawa R,Ames JB,Lagarias JC,Larsen DS

    更新日期:2021-01-13 00:00:00

  • Intramolecular triple-helix formation at (PunPyn).(PunPyn) tracts: recognition of alternate strands via Pu.PuPy and Py.PuPy base triplets.

    abstract::Triple-helical DNA shows increasing potential for applications in the control of gene expression (including therapeutics) and the development of sequence-specific DNA-cleaving agents. The major limitation in this technology has been the requirement of homopurine sequences for triplex formation. We describe a simple ap...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00117a002

    authors: Jayasena SD,Johnston BH

    更新日期:1992-01-21 00:00:00

  • Identification of tyrosine residues on ELMO1 that are phosphorylated by the Src-family kinase Hck.

    abstract::The SH3 and SH2 domains of hematopoietic cell kinase (Hck) play important roles in substrate targeting. To identify new components of Hck signaling pathways, we identified proteins that bind to the SH3 domain of Hck (Scott et al. (2002) J. Biol. Chem. 277, 28238). One such protein was ELMO1, the mammalian orthologue o...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi0500832

    authors: Yokoyama N,deBakker CD,Zappacosta F,Huddleston MJ,Annan RS,Ravichandran KS,Miller WT

    更新日期:2005-06-21 00:00:00

  • Internal dynamics of the tryptophan repressor (TrpR) and two functionally distinct TrpR variants, L75F-TrpR and A77V-TrpR, in their l-Trp-bound forms.

    abstract::Backbone amide dynamics of the Escherichia coli tryptophan repressor protein (WT-TrpR) and two functionally distinct variants, L75F-TrpR and A77V-TrpR, in their holo (l-tryptophan corepressor-bound) form have been characterized using (15)N nuclear magnetic resonance (NMR) relaxation. The three proteins possess very si...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi200389k

    authors: Tripet BP,Goel A,Copie V

    更新日期:2011-06-14 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...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00203a027

    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

  • 1H nuclear magnetic resonance double resonance study of oxytocin in aqueous solution.

    abstract::Peptide NH resonances in the 250 MHZ 1H nuclear magnetic resonance (NMR) spectrum of oxytocin in H2O were assigned to specific amino acid residues by the "underwater decoupling" technique (i.e., decoupling from corresponding CalphaH resonances, which are buried beneath the intense water peak). These experiments confir...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00650a025

    authors: Glickson JD,Rowan R,Pitner TP,Dadok J,Bothner-By AA,Walter R

    更新日期:1976-03-09 00:00:00

  • PRL1 promotes cell migration and invasion by increasing MMP2 and MMP9 expression through Src and ERK1/2 pathways.

    abstract::The PRL (phosphatase of regenerating liver) phosphatases represent a distinct class of protein tyrosine phosphatases, which are implicated in tumorigenesis and metastasis processes. Accumulating evidence indicates that alteration of PRL1 expression affects cell motility and tumor metastasis, although the biochemical p...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi8020789

    authors: Luo Y,Liang F,Zhang ZY

    更新日期:2009-03-03 00:00:00

  • Analysis of the AU-rich elements in the 3'-untranslated region of beta 2-adrenergic receptor mRNA by mutagenesis and identification of the homologous AU-rich region from different species.

    abstract::The 35000-Mr beta-adrenergic receptor mRNA binding protein (beta ARB) is induced by beta-adrenergic agonists and binds to G-protein-linked receptor mRNAs that exhibit agonist-induced destabilization. Recently, we identified a 20-nucleotide, AU-rich region in the 3'-untranslated region of the hamster beta 2-adrenergic ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9913348

    authors: Tholanikunnel BG,Raymond JR,Malbon CC

    更新日期:1999-11-23 00:00:00

  • Chemistry Is Dead. Long Live Chemistry!

    abstract::Chemistry, once king of fluorescence microscopy, was usurped by the field of fluorescent proteins. The increased demands of modern microscopy techniques on the "photon budget" require better and brighter fluorophores, causing a renewed interest in synthetic dyes. Here, we review the recent advances in biochemistry, pr...

    journal_title:Biochemistry

    pub_type: 杂志文章,评审

    doi:10.1021/acs.biochem.7b00529

    authors: Lavis LD

    更新日期:2017-10-03 00:00:00

  • Properties of the high-affinity single-stranded DNA binding state of the Escherichia coli recA protein.

    abstract::The properties of the high-affinity single-stranded DNA (ssDNA) binding state of Escherichia coli recA protein have been studied. We find that all of the nucleoside triphosphates that are hydrolyzed by recA protein induce a high-affinity ssDNA binding state. The effect of ATP binding to recA protein was partially sepa...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00404a021

    authors: Menetski JP,Varghese A,Kowalczykowski SC

    更新日期:1988-02-23 00:00:00

  • Atomic force microscopy imaging reveals the domain structure of polycystin-1.

    abstract::Mutation of polycystin-1 (PC1) is the major cause of autosomal dominant polycystic kidney disease. PC1 has a predicted molecular mass of ~460 kDa comprising a long multidomain extracellular N-terminal region, 11 transmembrane regions, and a short C-terminal region. Because of its size, PC1 has proven difficult to hand...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi300134b

    authors: Oatley P,Stewart AP,Sandford R,Edwardson JM

    更新日期:2012-04-03 00:00:00

  • Evidence for a methylammonium-binding site on methylamine dehydrogenase of Thiobacillus versutus.

    abstract::The nonconvertible substrate analogues di-, tri-, and tetramethylammonium are bound with fairly high affinity to oxidized methylamine dehydrogenase (MADHox) from Thiobacillus versutus and induce the same red-shift in the optical absorbance spectrum of MADHox as do the monovalent cations Cs+, Rb+, and NH4+. Like the mo...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00040a002

    authors: Gorren AC,Moenne-Loccoz P,Backes G,de Vries S,Sanders-Loehr J,Duine JA

    更新日期:1995-10-10 00:00:00

  • Molecular dynamics simulations of the TrkH membrane protein.

    abstract::TrkH is a transmembrane protein that mediates uptake of K(+) through the cell membrane. Despite the recent determination of its crystallographic structure, the nature of the permeation mechanism is still unknown, that is, whether K(+) ions move across TrkH by active transport or passive diffusion. Here, molecular dyna...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi201586n

    authors: Domene C,Furini S

    更新日期:2012-02-28 00:00:00

  • A second photochromic bacteriophytochrome from Synechocystis sp. PCC 6803: spectral analysis and down-regulation by light.

    abstract::It now appears that photosynthetic prokaryotes and lower eukaryotes possess higher plant phytochrome-like proteins. In this work, a second phytochrome-like gene was isolated, in addition to the recently identified Cph1 phytochrome, from the Synechocystis sp. PCC 6803, and its gene product was characterized photochemic...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi992831r

    authors: Park CM,Kim JI,Yang SS,Kang JG,Kang JH,Shim JY,Chung YH,Park YM,Song PS

    更新日期:2000-09-05 00:00:00

  • Heme-CO religation in photolyzed hemoglobin: a time-resolved Raman study of the Fe-CO stretching mode.

    abstract::Time-resolved resonance Raman spectroscopy has been employed to monitor geminate heme-CO rebinding in photolyzed HbCO. The excitation frequency was tuned to enhance the scattering from rebound heme sites 20-500 ns subsequent to CO photolysis. The behavior of vFe-C during ligand rebinding has important ramifications co...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00056a017

    authors: Schneebeck MC,Vigil LE,Friedman JM,Chavez MD,Ondrias MR

    更新日期:1993-02-09 00:00:00

  • Optical spectroscopic characterization of single tryptophan mutants of chicken skeletal troponin C: evidence for interdomain interaction.

    abstract::The effects of metal ion binding on the optical spectroscopic properties and temperature stability of two single tryptophan mutants of chicken skeletal TnC, F78W and F154W, have been examined. The absence of tyrosine and other tryptophan residues allowed the unambiguous assignment of the spectral signal from the intro...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi982048j

    authors: Moncrieffe MC,Venyaminov SY,Miller TE,Guzman G,Potter JD,Prendergast FG

    更新日期:1999-09-14 00:00:00

  • Mechanism of enolase: the crystal structure of enolase-Mg2(+)-2-phosphoglycerate/phosphoenolpyruvate complex at 2.2-A resolution.

    abstract::Enolase in the presence of Mg2+ catalyzes the elimination of H2O from 2-phosphoglyceric acid (PGA) to form phosphoenolpyruvate (PEP) and the reverse reaction, the hydration of PEP to PGA. The structure of the ternary complex yeast enolase-Mg2(+)-PGA/PEP has been determined by X-ray diffraction and refined by crystallo...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00225a012

    authors: Lebioda L,Stec B

    更新日期:1991-03-19 00:00:00

  • Slow-binding inhibition of Escherichia coli cystathionine beta-lyase by L-aminoethoxyvinylglycine: a kinetic and X-ray study.

    abstract::The pyridoxal 5'-phosphate (PLP)-dependent cystathionine beta-lyase (CBL) was previously found to be inhibited by the natural toxins rhizobitoxine and l-aminoethoxyvinylglycine (AVG). The present study characterizes the interaction of Escherichia coli CBL with AVG and methoxyvinylglycine (MVG) by a combination of kine...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi970630m

    authors: Clausen T,Huber R,Messerschmidt A,Pohlenz HD,Laber B

    更新日期:1997-10-14 00:00:00

  • Competing Protonation and Halide Elimination as a Probe of the Character of Thiamin-Derived Reactive Intermediates.

    abstract::Decarboxylation reactions from comparable thiamin diphosphate- and thiamin-derived adducts of p-(halomethyl)benzoylformic acids in enzymic and non-enzymic reactions, respectively, reveal critical distinctions in otherwise similar Breslow intermediates. The ratio of protonation to chloride elimination from the Breslow ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.9b00298

    authors: Bielecki M,Howe GW,Kluger R

    更新日期:2019-08-27 00:00:00

  • Analysis of neuronal NO synthase under single-turnover conditions: conversion of Nomega-hydroxyarginine to nitric oxide and citrulline.

    abstract::Nitric oxide synthases (NOSs) are proposed to generate NO and citrulline from L-arginine in two steps: initial N-hydroxylation to generate Nomega-hydroxyarginine (NOHA) followed by a three-electron oxidation of the hydroxylated nitrogen to form products. Both steps consume NADPH and may involve heme iron-based activat...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi971414g

    authors: Abu-Soud HM,Presta A,Mayer B,Stuehr DJ

    更新日期:1997-09-09 00:00:00

  • Design of the helix-turn-helix motif: nonlocal effects of quaternary structure in DNA recognition investigated by laser Raman spectroscopy.

    abstract::The operator-binding domain of phage lambda repressor provides a model for DNA recognition by the helix-turn-helix (HTH) motif. In the wild-type protein, dimerization is mediated by hydrophobic packing (of the dyad-related helix 5), which serves as an indirect determinant of operator affinity. The mutant repressor, Ty...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00232a003

    authors: Benevides JM,Weiss MA,Thomas GJ Jr

    更新日期:1991-05-07 00:00:00

  • Secondary structure in properdin of the complement cascade and related proteins: a study by Fourier transform infrared spectroscopy.

    abstract::Six structural repeat motifs of 58 amino acids are found in the sequence of both mouse and human properdins. Twelve more examples of the motif are available from the sequences of thrombospondin, the terminal complement components, and the thrombospondin-related anonymous protein. The averaged Robson and Chou-Fasman se...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00444a007

    authors: Perkins SJ,Nealis AS,Haris PI,Chapman D,Goundis D,Reid KB

    更新日期:1989-09-05 00:00:00

  • The amino acid sequence of a carboxypeptidase inhibitor from potatoes.

    abstract::The carboxypeptidase inhibitor from Russet Burbank potatoes (C. A. Ryan et al. (1974b), J. Biol. Chem 249, 5495) is a mixture of approximately equal amounts of two polypeptide chains containing 38 and 39 amino acid residues, respectively. The chains differ in their amino terminal sequence only, one beginning with smal...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00677a036

    authors: Hass GM,Nau H,Biemann K,Grahn DT,Ericsson LH,Neurath H

    更新日期:1975-03-25 00:00:00

  • Use of a photoactivated ruthenium dimer complex to measure electron transfer between the Rieske iron-sulfur protein and cytochrome c(1) in the cytochrome bc(1) complex.

    abstract::Electron transfer between the Rieske iron-sulfur protein (Fe(2)S(2)) and cytochrome c(1) was studied using the ruthenium dimer, Ru(2)D, to either photoreduce or photooxidize cytochrome c(1) within 1 micros. Ru(2)D has a charge of +4, which allows it to bind with high affinity to the cytochrome bc(1) complex. Flash pho...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi000003o

    authors: Sadoski RC,Engstrom G,Tian H,Zhang L,Yu CA,Yu L,Durham B,Millett F

    更新日期:2000-04-18 00:00:00

  • Oxidative titrations of reduced cytochrome aa3: anisotropic extinction behavior observed in the heme alpha-band region.

    abstract::Direct chemical titrations of reduced, purified cytochrome aa3 were monitored at 604 nm. Anaerobic oxidation by potassium ferricyanide or 1,1'-bis(hydroxymethyl)-ferricinium ion was compared with the natural substrate, molecular oxygen. The four electrons from reduced cytochrome aa3 were donated to one oxygen molecule...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00626a014

    authors: Schroedl NA,Hartzell CR

    更新日期:1977-04-05 00:00:00