Abstract:
:Bacterial bioluminescence is a phenomenon resulting from the reaction of a two-component FMN-dependent aldehyde monooxygenase system, which comprises a bacterial luciferase and a flavin reductase. Bacterial luciferase (LuxAB) is one of the most extensively investigated two-component monooxygenases, while its reductase partner, the flavin reductase (LuxG) from the same operon, has only been recently expressed in a functional form. This work reports transient kinetics identification of intermediates in the LuxG reaction using stopped-flow spectrophotometry. The results indicate that the overall reaction follows a sequential-ordered mechanism in which NADH binds first to the enzyme, followed by FMN, resulting in the formation of charge-transfer intermediate 1 (CT-1) typical of those between reduced pyridine nucleotides and oxidized flavins. The next step is the reduction of FMN as indicated by a large decrease in absorbance at 450 nm. The reduction of FMN is biphasic. The first phase of FMN reduction occurs concurrently with formation of charge-transfer intermediate 2 (CT-2), while the second phase is synchronous with the decay of CT-2. When the isotope-labeled substrate, 4(R)-[(2)H]NADH, was used, the first reduction phase showed a primary kinetic isotope effect ((D)k(red)) of ≥3.9 and resulted in greater accumulation of CT-1. These results are consistent with CT-1 being the FMN(ox):NADH complex, while CT-2 is the FMN(red):NAD(+) complex. Because CT-2 decays with a rate constant of 2.8 ± 0.2 s(-1), while the turnover number obtained from the steady-steady-state kinetics is 1.7 s(-1), it is likely that the CT-2 decay step largely controls the overall reaction rate. All kinetic data are consistent with a half-sites reactivity model in which flavin reduction occurs at only one subunit at a time. The first reduction phase is due to the reduction of FMN in the first subunit, while the second phase is due to the reduction of FMN in the second subunit. The latter phase is limited by the rate of decay of CT-2 in the first subunit. The half-sites reactivity model is also supported by detection of burst kinetics during the pre-steady-state period that is correlated with 0.5 mol of the FMN being reduced/mol of the LuxG:NADH complex. The functional importance of this half-site reactivity phenomenon is still unclear.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Nijvipakul S,Ballou DP,Chaiyen Pdoi
10.1021/bi1009985subject
Has Abstractpub_date
2010-11-02 00:00:00pages
9241-8issue
43eissn
0006-2960issn
1520-4995journal_volume
49pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The plasma membrane of the Ehrlich ascites tumor cell contains an NADH dehydrogenase. This activity was shown not to be due to contamination by other subcellular membranes. A variety of electron acceptors have been compared as to rate with the following result: ferricyanide greater than cytochrome c greater than cytoc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00575a021
更新日期:1979-04-17 00:00:00
abstract::The acid-base properties of ionophore A23187 in methanol-water solutions (0--95% w/w) and bound to unilamellar vesicles of dimyristoylphosphatidylcholine were examined by ultraviolet and fluorescence spectroscopy, and the spectral properties for the acidic and basic forms were defined under these conditions. Standard ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00539a023
更新日期:1982-05-11 00:00:00
abstract::Chloroperoxidase is a versatile fungal heme-thiolate protein that catalyzes a variety of one-electron and two-electron oxidations. We report here that the alkylation of an essential histidine residue showed no effect on the one-electron peroxidations but inhibited two-electron oxidations. The pH profiles of different ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7022656
更新日期:2008-03-04 00:00:00
abstract::Post-translational phosphorylation and the related conformational changes in signaling proteins are responsible for regulating a wide range of subcellular processes. Human Pin1 is central to many of these cell signaling pathways in normal and aberrant subcellular processes, catalyzing cis-trans isomerization of the pe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi2009954
更新日期:2011-11-08 00:00:00
abstract::To investigate the effect of a reduced level of selenocysteine (Sec) tRNA[Ser]Sec in selenoprotein biosynthesis, two mouse embryonic stem (ES) cell lines heterozygous for the corresponding gene were generated by homologous recombination of the host genome with targeting vectors encoding a deleted or a disrupted tRNA[S...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970608t
更新日期:1997-07-15 00:00:00
abstract::The family 4 uracil-DNA glycosylase from the hyperthermophilic organism Archaeoglobus fulgidus (AFUDG) is responsible for the removal of uracil in DNA as the first step in the base excision repair (BER) pathway. AFUDG contains a large solvent-exposed peptide region containing an α helix and loop anchored on each end v...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi3000462
更新日期:2012-06-26 00:00:00
abstract::The unique characteristics of DNA charge transport (CT) have prompted an examination of roles for this chemistry within a biological context. Not only can DNA CT facilitate long-range oxidative damage of DNA, but redox-active proteins can couple to the DNA base stack and participate in long-range redox reactions using...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/bi501520w
更新日期:2015-02-03 00:00:00
abstract::The cAMP-dependent protein kinase (cAPK) plays an essential role during differentiation and fruit morphogenesis in Dictyostelium discoideum. The presence of an open reading frame on the gene, pkaC (previously named either Dd PK2 or Dd PK3 by different groups), predicts a 73-kDa polypeptide with 54% similarity to the c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00088a003
更新日期:1993-09-21 00:00:00
abstract::Factor V and factor Va binding to single bilayer phospholipid vesicles was investigated by light-scattering intensity measurements. This technique allows the measurement of free and phospholipid-bound protein concentrations from which equilibrium constants can be obtained. As controls, the Ca2+-dependent phospholipid ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00587a023
更新日期:1979-10-02 00:00:00
abstract::Folding experiments have suggested that some proteins have kinetic intermediates with a non-native structure. A simple G ̅o model does not explain such non-native intermediates. Therefore, the folding energy landscape of proteins with non-native intermediates should have characteristic properties. To identify such pro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi200241a
更新日期:2011-07-26 00:00:00
abstract::The effect of long-term storage on the electronic absorption spectrum and the kinetic properties of cytochrome c peroxidase has been investigated. No detectable differences were observed between freshly isolated enzyme and enzyme stored below -20 degrees C, in the crystalline state, for up to 41 months. The electronic...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00470a004
更新日期:1990-05-08 00:00:00
abstract::Amyloid diseases make up a set of fatal disorders in which proteins aggregate to form fibrils that deposit in tissues throughout the body. Amyloid-associated diseases are challenging to study because amyloid formation occurs on time scales that span several orders of magnitude and involve heterogeneous, interconvertin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.9b00769
更新日期:2019-11-05 00:00:00
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...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00445a004
更新日期:1989-09-19 00:00:00
abstract::Previously we showed that the low-molecular-weight mucin (MG2, encoded by MUC7), a major component of human submandibular/sublingual saliva, is a bacterial receptor that coats the tooth surface. Here we tested the hypothesis that the structure of its carbohydrate residues contains important information about its funct...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi972612a
更新日期:1998-04-07 00:00:00
abstract::West Nile virus (WNV) is a mosquito-borne flavivirus with a rapidly expanding global distribution. Infection can cause severe neurological disease and fatality in humans. Efforts are ongoing to develop antiviral drugs that inhibit the WNV protease, a viral enzyme required for polyprotein processing. Unfortunately, lit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi400782w
更新日期:2013-10-01 00:00:00
abstract::Some properties of [4-vinyl] chlorophyllide a reductase are described. This enzyme converts divinyl chlorophyllide a to monovinyl chlorophyllide a. The latter is the immediate precursor of monovinyl chlorophyll a, the main chlorophyll in green plants. [4-Vinyl] chlorophyllide a reductase plays an important role in day...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00151a011
更新日期:1992-09-15 00:00:00
abstract::By superimposing data obtained by photo-cross-linking RGD-containing ligands to the human alpha(V)beta(3) integrin onto the crystal structure of the ectopic domain of this receptor (Xiong et al. (2001) Science 294, 339-345), we have identified the binding site for the RGD triad within this integrin. We synthesized thr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi025690t
更新日期:2002-07-02 00:00:00
abstract::The mitochondrial folate transporter (MFT) was previously identified in human and hamster cells. Sequence homology of this protein with the inner mitochondrial membrane transporters suggested a domain structure in which the N- and C-termini of the protein are located on the mitochondrial intermembrane-facing surface, ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi062191+
更新日期:2007-02-13 00:00:00
abstract::Two photoreactive apamin derivatives were prepared with an aryl azide [[(azidonitrophenyl)amino]acetate (ANPAA)] group coupled at different positions on the neurotoxin molecule. These ligands were used to identify membrane components in the environment of the neuronal binding site that is associated with a Ca2+-activa...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00362a010
更新日期:1986-07-15 00:00:00
abstract::Vibrational frequencies associated with FeC and CO stretching and FeCO bending modes have been determined via resonance Raman (RR) and infrared (IR) spectroscopy for cytochrome c peroxidase (CCP) mutants prepared by site-directed mutagenesis. These include the bacterial "wild type", CCP(MI), and mutations involving gr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00415a015
更新日期:1988-07-26 00:00:00
abstract::Activation of washed human platelets initiated with alpha-thrombin, SFLLRN, or AYPGKF invariably results in the generation of PAR-1-(1-41) and PAR-4-(1-47). PAR-1-(1-41) and PAR-4-(1-47) are amino-terminal peptides generated when PAR-1 and -4 are cleaved in their first extracellular domains after R(41) and R(47), resp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi801399c
更新日期:2009-02-24 00:00:00
abstract::Illuminating Ca2+-depleted photosystem (PS) II membranes generated two new EPR signals at g = 11 and 15 by perpendicular and parallel polarization modes, respectively. Two turnovers of the oxygen evolving center (OEC) beyond the modified S2' state are required for the appearance of these signals. The formation of the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi972471c
更新日期:1998-03-03 00:00:00
abstract::Over 155 mutations within the V2 vasopressin receptor (AVPR2) gene are responsible for nephrogenic diabetes insipidus (NDI). The expression and subcellular distribution of four of these was investigated in transfected cells. These include a point mutation in the seventh transmembrane domain (S315R), a frameshift mutat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002699r
更新日期:2001-06-12 00:00:00
abstract::ABCG2 is a half-ATP binding cassette (ABC) drug transporter that consists of a nucleotide binding domain (NBD) followed by a transmembrane domain. This half-ABC transporter is thought to form a homodimer in the plasma membrane where it transports anticancer drugs across the biological membranes in an ATP-dependent man...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi900675v
更新日期:2009-09-29 00:00:00
abstract::Drug efflux and enzymatic drug degradation are two cellular mechanisms that contribute to drug resistance in many cancers. Herein, we report the synthesis and in vitro activity of a pro-immunostimulant that exploits both processes in tandem to selectively confer cancer-mediated immunogenicity. We demonstrate that an i...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00095
更新日期:2018-04-17 00:00:00
abstract::The metalloregulatory protein MntR from Bacillus subtilis acts as a transcriptional regulator of manganese homeostasis. MntR is a member of a subfamily of DtxR-related proteins that perform analogous regulatory functions in a variety of pathogenic organisms. Metal ions activate MntR to bind DNA and repress the transcr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0350248
更新日期:2003-11-04 00:00:00
abstract::The X-ray crystal structure of recombinant Desulfovibrio vulgaris rubrerythrin (Rbr) that was subjected to metal constitution first with zinc and then iron, yielding ZnS(4)Rbr, is reported. A [Zn(SCys)(4)] site with no iron and a diiron site with no appreciable zinc in ZnS(4)Rbr were confirmed by analysis of the anoma...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0356193
更新日期:2004-03-23 00:00:00
abstract::Ligand-induced dimerization of receptor tyrosine kinases (RTKs) modulates a system of linked biochemical reactions, sharply switching the RTK from a quiescent state to an active state that becomes phosphorylated and triggers intracellular signaling pathways. To improve our understanding of this molecular switch, we de...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi701892f
更新日期:2008-04-01 00:00:00
abstract::Aqueous suspensions of dark-adapted bacteriorhodopsin (bR560) in the purple membrane of Halobacterium halobium are exposed to rapid jumps to high pH. Optical and resonance Raman measurements are carried out by using flow and stationary methods. Above pH congruent to 11.5 bR560 starts to be reversibly converted to a sp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00263a019
更新日期:1982-09-28 00:00:00
abstract::Treatment of cells with the enediyne C-1027 is highly efficient at inducing single- and double-strand DNA breaks. This agent is highly cytotoxic when used at picomolar levels over a period of days. For this study, C-1027 has been used at higher levels for a much shorter time period to look at early cellular responses ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi015680c
更新日期:2001-12-04 00:00:00