Abstract:
:The steady-state tryptophan fluorescence and time-resolved tryptophan fluorescence of Escherichia coli thioredoxin, calf thymus thioredoxin, and yeast thioredoxin have been studied. In all proteins, the tryptophan residues undergo strong static and dynamic quenching, probably due to charge-transfer interactions with the nearby sulfur atoms of the active cysteines. The use of a high-resolution photon counting instrument, with a time response of 60 ps full width at half-maximum, allowed the detection of fluorescence lifetimes ranging from a few tens of picoseconds to 10 ns. The data were analyzed both by classical nonlinear least squares and by a new method of entropy maximization (MEM) for the recovery of lifetime distributions. Simulations representative of the experimental data were used to test the MEM analysis. Strong support was obtained in this way for a small number of averaged discrete species in the fluorescence decays. Wavelength studies show that each of these components spreads over closely spaced excited states, while the temperature studies indicate that they do not exchange significantly on the nanosecond time scale. The oxidized form of thioredoxin is characterized by a high content of a very short lifetime below 70 ps, the amplitude of which is sharply decreased upon reduction. On the other hand, the fluorescence anisotropy decays indicate that reduction causes an increase of the very fast tryptophan rotations in an otherwise relatively rigid structure. While the calf thymus and E. coli proteins have mostly similar dynamical fluorescence properties, the yeast thioredoxin differs in many respects.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Mérola F,Rigler R,Holmgren A,Brochon JCdoi
10.1021/bi00434a038subject
Has Abstractpub_date
1989-04-18 00:00:00pages
3383-98issue
8eissn
0006-2960issn
1520-4995journal_volume
28pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Heparanase is a beta-D-endoglucuronidase that cleaves heparan sulfate (HS) and has been implicated in many important physiological and pathological processes, including tumor cell metastasis, angiogenesis, and leukocyte migration. We report herein the identification of active-site residues of human heparanase. Using P...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002080p
更新日期:2000-12-26 00:00:00
abstract::Folding models suggest that the highly conserved histidine 217 of the cytochrome b subunit from the cytochrome bc1 complex is close to the quinone reductase (Qi) site. This histidine (bH217) in the cytochrome b polypeptide of the photosynthetic bacterium Rhodobacter capsulatus has been replaced with three other residu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00169a014
更新日期:1994-01-25 00:00:00
abstract::To better understand origin recognition and initiation of DNA replication, we have examined by NMR complexes formed between the origin-binding domain of SV40 T antigen (T-ag-obd), the initiator protein of the SV40 virus, and cognate and noncognate DNA oligomers. The results reveal two structural effects associated wit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi030228+
更新日期:2004-06-08 00:00:00
abstract::c-Myc is a member of a family of sequence specific-DNA binding proteins that are thought to regulate the transcription of genes involved in normal cell growth, differentiation, and apoptosis. In order to understand how human c-myc functions as a transcription factor, we have studied the mechanism of action and structu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960793v
更新日期:1996-07-23 00:00:00
abstract::Treatment of soybean lipoxygenase isozyme 1 with its substrates, linoleic acid and oxygen, or product, 13(S)-hydroperoxy-9,11(Z,E)-octadecadienoic acid (13-HPOD), leads to the appearance of a purple color. Although the structure of the chromophore has not been determined, we present strong evidence that it is an Fe(3+...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00018a019
更新日期:1995-05-09 00:00:00
abstract::Photoreceptor guanylyl cyclase (ROS-GC), converting GTP into cGMP and pyrophosphate, is a key enzyme in the regulation of the visual transduction cascade. ROS-GC requires GC-activating proteins (GCAPs) and low free [Ca] for full activity. We found that when choline or potassium were the major cations present, light ca...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9825596
更新日期:1999-03-02 00:00:00
abstract::Neutral solutes were used to investigate the effects of osmotic stress both on the ability of rhodopsin to undergo its activating conformation change and on acyl chain packing in the rod outer segment (ROS) disk membrane. The equilibrium concentration of metarhodopsin II (MII), the conformation of photoactivated rhodo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990634m
更新日期:1999-06-15 00:00:00
abstract::NADPH-cytochrome P450 oxidoreductase (P450 reductase, EC 1.6.2.4) is an essential component of the P450 monooxygenase complex and binds FMN, FAD, and NADPH cofactors. Residues Tyr140 and Tyr178 are known to be involved in FMN binding. A third aromatic side chain, Phe181, is also located in the proximity of the FMN rin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi011147l
更新日期:2001-11-13 00:00:00
abstract::Ppr from the purple phototrophic bacterium, Rhodospirillum centenum (also known as Rhodocista centenaria), is a hybrid of photoactive yellow protein (PYP), bacteriophytochrome (Bph), and histidine kinase (HK) domains. The holo-Ppr (containing both chromophores) exhibits characteristic absorption maxima at 435 nm due t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700616j
更新日期:2007-07-17 00:00:00
abstract::Primary kinetic isotope effects on Vmax were observed in both reaction directions upon racemizing samples of [2-2H]glutamate with the cofactor-independent glutamate racemase from Lactobacillus. This supports a deprotonation/protonation mechanism for racemization in which the breaking of the carbon-hydrogen bond at C-2...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00066a021
更新日期:1993-04-20 00:00:00
abstract::The interaction of insulin with its receptor is complex. Kinetic and equilibrium binding studies suggest coexistence of high- and low-affinity binding sites or negative cooperativity. These phenomena and high-affinity interactions are dependent on the dimeric structure of the receptor. Structure-function studies of in...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi801693h
更新日期:2008-12-02 00:00:00
abstract::Vitamin B12-dependent methionine synthase catalyzes the transfer of a methyl group from methyltetrahydrofolate to homocysteine via the enzyme-bound cofactor methylcobalamin. To carry out this reaction, the enzyme must alternately stabilize six-coordinate methylcobalamin and four-coordinate cob(I)alamin oxidation state...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952389m
更新日期:1996-02-20 00:00:00
abstract::DADH catalyzes the flavin-dependent oxidative deamination of d-amino acids to the corresponding α-keto acids and ammonia. Here we report the first X-ray crystal structures of DADH at 1.06 Å resolution and its complexes with iminoarginine (DADH(red)/iminoarginine) and iminohistidine (DADH(red)/iminohistidine) at 1.30 Å...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1005865
更新日期:2010-10-05 00:00:00
abstract::We describe the crystal structure of human epidermal-type fatty acid binding protein (E-FABP) that was recently found to be highly upregulated in human psoriatic keratinocytes. To characterize E-FABP with respect to ligand-binding properties and tertiary structure, we cloned the respective cDNA, overexpressed the prot...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990305u
更新日期:1999-09-21 00:00:00
abstract::Life span regulation and inhibition of gene silencing in yeast have been linked to nicotinamide effects on Sir2 enzymes. The Sir2 enzymes are NAD(+)-dependent protein deacetylases that influence gene expression by forming deacetylated proteins, nicotinamide and 2'-O-acetyl-ADPR. Nicotinamide is a base-exchange substra...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi034959l
更新日期:2003-08-12 00:00:00
abstract::The far-ultraviolet circular dichroism spectra of bovine and human prothrombin, prothrombin fragment 1, prethrombin 1, prothrombin fragment 2, and prethrombin 2 (prethrombin 2des(1--13)) were determined and the method of Chen et al. [Chen, Y. H., Yang, J. T., & Martinez, H. M. (1972) Biochemistry 11, 4120--4131; Chen,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00577a017
更新日期:1979-05-15 00:00:00
abstract::Two transcription factors, CRP and CytR, mediate positive and negative control of nine cistrons involved in nucleoside catabolism and recycling in Escherichia coli. The ability of multiple transcription factors to combine in different ways to confer differential gene regulation is of significant interest in both proka...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901583n
更新日期:2010-01-26 00:00:00
abstract::Over 70 transthyretin (TTR) mutations facilitate amyloidosis in tissues other than the central nervous system (CNS). In contrast, the D18G TTR mutation in individuals of Hungarian descent leads to CNS amyloidosis. D18G forms inclusion bodies in Escherichia coli, unlike the other disease-associated TTR variants overexp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi027319b
更新日期:2003-06-10 00:00:00
abstract::The first positive evidence for the utilization of a direct C-6' ' oxidation/reduction mechanism by ADP-l-glycero-d-manno-heptose 6-epimerase is reported here. The epimerase (HldD or AGME, formerly RfaD) operates in the biosynthetic pathway of l-glycero-d-manno-heptose, which is a conserved sugar in the core region of...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi050106c
更新日期:2005-04-19 00:00:00
abstract::The conformation and activity of three different lipases have been studied in reverse micelles formed by sodium bis(2-ethylhexyl) sulfosuccinate (AOT) in isooctane. In the case of human pancreatic lipase, the conformation of the polypeptide chain--as judged from far-UV circular dichroism measurements--is only slightly...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00066a025
更新日期:1993-04-20 00:00:00
abstract::Methylenetetrahydrofolate dehydrogenase-cyclohydrolase (FolD) catalyzes interconversion of 5,10-methylene-tetrahydrofolate and 10-formyl-tetrahydrofolate in the one-carbon metabolic pathway. In some organisms, the essential requirement of 10-formyl-tetrahydrofolate may also be fulfilled by formyltetrahydrofolate synth...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00400
更新日期:2015-06-09 00:00:00
abstract::Myosin belongs to the family of motor proteins. Its interaction with actin coupled with hydrolysis of ATP is the molecular basis of muscle contraction. The head segment of myosin, called subfragment 1 (S1), contains the distinct binding sites for ATP and actin and responsible for the ATPase activity. The rate-limiting...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952565r
更新日期:1996-04-09 00:00:00
abstract::The binding of the benzo[a]pyrene metabolite anti-BPDE (r7, t8-dihydroxy-t9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene) to the N(2) group of 2'-deoxyguanosine residues (dG) is known to adversely affect the Michaelis-Menten primer extension kinetics catalyzed by DNA Pol I and other polymerases. In this work, the impact...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990614k
更新日期:1999-09-07 00:00:00
abstract::DB2277, a heterocyclic diamidine, is a successful design for mixed base pair (bp) DNA sequence recognition. The compound has a central aza-benzimidazole group that forms two H-bonds with a GC bp that has flanking AT bps. The nuclear magnetic resonance structure of the DB2277-DNA complex with an AAGATA recognition site...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00647
更新日期:2018-08-21 00:00:00
abstract::The inducible isoform of nitric oxide synthase (iNOS) and three zinc tetrathiolate mutants (C104A, C109A, and C104A/C109A) were expressed in Escherichia coli and purified. The mutants were found by ICP-AES and the zinc-specific PAR colorimetric assay to be zinc free, whereas the wild-type iNOS zinc content was 0.38 +/...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0474463
更新日期:2005-03-29 00:00:00
abstract::We have developed a new genomic sequencing method for detecting, with resolution at the nucleotide level, the interstrand DNA cross-links induced by 4,5',8-trimethylpsoralen along single-copy genes in mammalian cells. The cross-links (diadducts) initially formed are converted into monoadducts by alkali reversal prior ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002539f
更新日期:2001-04-03 00:00:00
abstract::Hemoglobins extracted from fishes that live in temperate waters show little or no dissociation even in the liganded form, unlike human hemoglobin (HbA). To establish whether cold adaptation influences the tendency to dissociate, the dimer-tetramer association constants (L(2,4)) of the carbonmonoxy derivatives of repre...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002297j
更新日期:2001-03-13 00:00:00
abstract::The mutagenic and genotoxic properties of 1,N6-ethenoadenine (epsilon Ade), 3,N4-ethenocytosine (epsilon Cyt), and 4-amino-5-(imidazol-2-yl)imidazole (beta) were investigated in vivo. The former two modified bases are known DNA adducts formed by the human carcinogen vinyl chloride; beta is formed by pyrimidine ring-op...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00210a031
更新日期:1993-11-30 00:00:00
abstract::The reduction of D-amino acid oxidase (DAAO) by hydrated electrons (eaq-) has been studied in the absence and presence of benzoate by pulse radiolysis. The eaq-did not reduce the flavin moiety in DAAO and reacted with the amino acid residues in the protein. In the presence of benzoate, eaq- first reacted with benzoate...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00278a028
更新日期:1983-04-26 00:00:00
abstract::The specificity of trypsin for Arg- and Lys-containing substrates depends upon the presence of Asp189 at the base of the primary binding pocket. The crystal structure of anionic rat trypsin D189S complexed with BPTI reveals that removal of the aspartate side chain permits the binding of a well-ordered acetate ion in a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00177a016
更新日期:1994-03-22 00:00:00