Abstract:
:Excluded volume and viscosity effects of crowding agents that mimic crowded conditions in vivo on "classical" burst phase folding kinetics of cytochrome c are assessed in vitro. Upon electron transfer-triggered folding of reduced cytochrome c, far-UV time-resolved circular dichroism (TRCD) is used to monitor folding under different conditions. Earlier work has shown that folding of reduced cytochrome c from the guanidinium hydrochloride-induced unfolded ensemble in dilute phosphate buffer involves kinetic partitioning: one fraction of molecules folds rapidly, on a time scale identical to that of reduction, while the remaining population folds more slowly. In the presence of 220 mg/mL dextran 70, a synthetic macromolecular crowding agent that occupies space but does not interact with proteins, the population of the fast folding step for cytochrome c is greatly reduced. Increasing the viscosity with sucrose to the same microviscosity exhibited by the dextran solution showed no significant decrease in the amplitude of the fast-folding phase of cytochrome c. Experiments show that the unfolded-state heme ligation remains bis-His in the presence of dextran 70, but coarse-grained simulations suggest that the unfolded-state ensemble becomes more compact in the presence of crowders. We conclude that excluded volume effects alter unfolded cytochrome c such that access to fast-folding conformations is reduced.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Chen E,Christiansen A,Wang Q,Cheung MS,Kliger DS,Wittung-Stafshede Pdoi
10.1021/bi301324ysubject
Has Abstractpub_date
2012-12-11 00:00:00pages
9836-45issue
49eissn
0006-2960issn
1520-4995journal_volume
51pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Protein enzymes often use ionizable side chains, such as histidine, for general acid-base catalysis because the imidazole pK(a) is near neutral pH. RNA enzymes, on the other hand, are comprised of nucleotides which do not have apparent pK(a) values near neutral pH. Nevertheless, it has been recently shown that cytidin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi001976r
更新日期:2000-12-26 00:00:00
abstract::Band 3, the anion transport protein of human erythrocyte membranes, can be transferred from cells to liposomes and from liposomes back to cell membranes, retaining function and native orientation. After incubation with cells, sonicated phosphatidylcholine vesicles bind a transmembrane protein that comigrates with band...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00295a011
更新日期:1983-12-20 00:00:00
abstract::Retinal membrane guanylyl cyclase (RetGC) in the outer segments of vertebrate photoreceptors is controlled by guanylyl cyclase activating proteins (GCAPs), responding to light-dependent changes of the intracellular Ca(2+) concentrations. We present evidence that a different RetGC binding protein, retinal degeneration ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi201342b
更新日期:2011-11-08 00:00:00
abstract::Frequency-domain fluorescence resonance energy transfer and anisotropy measurements were performed to characterize conformational dynamics of an analog of the RNase S-peptide (residues 1-20). Trp was used as a donor by replacing Phe 8, and a dansyl acceptor group was introduced at position 1 or 18. The distance-distri...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00097a009
更新日期:1993-11-23 00:00:00
abstract::Aerobic and facultative bacteria and archaea harboring mer loci exhibit resistance to the toxic effects of Hg(II) and organomercurials [RHg(I)]. In broad spectrum resistance, RHg(I) is converted to less toxic Hg(0) in the cytosol by the sequential action of organomercurial lyase (MerB: RHg(I) → RH + Hg(II)) and mercur...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi100802k
更新日期:2010-09-21 00:00:00
abstract::Pyruvate decarboxylase (PDC) uses thiamine diphosphate as an essential cofactor to catalyze the formation of acetaldehyde on the pathway of ethanol synthesis. Here we report the crystallographic image of a prereaction intermediate of a bacterial pyruvate decarboxylase prepared by cocrystallizing the enzyme with pyruva...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901864j
更新日期:2010-03-02 00:00:00
abstract::We recently introduced a new substrate rescue tool for investigating enzymatic base flipping by uracil DNA glycosylase (UDG) in which a bulky pyrene nucleotide wedge (Y) was placed opposite a uracil in duplex DNA (i.e., a U/Y pair), thereby preorganizing the target base in an extrahelical conformation [Jiang, Y. L., e...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026227j
更新日期:2002-09-17 00:00:00
abstract::The structure/function properties of functional groups in the leadzyme have been studied by assaying the activity of analog ribozymes generated by the systematic substitution of modified nucleotides in the internal loop region of the ribozyme. Guanosine analogs introduced at positions 4 and 7 occupied by guanosine in ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962219p
更新日期:1997-03-18 00:00:00
abstract::We have measured the base-pair lifetimes in oligodeoxynucleotides containing tracts of A.T base pairs using imino proton magnetic resonance. When the tract contains more than four consecutive A.T base pairs, possibly including a 5'-AT step but not a 5'-TA step, anomalously long lifetimes are observed. For example, the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00425a004
更新日期:1988-12-13 00:00:00
abstract::To identify residues and segments in the central region of apolipoprotein A-I (apoA-I) that are important for the protein structure and stability, we studied the effects of four double charge ablations, D102A/D103A, E110A/E111A, R116V/K118A, and R160V/H162A, and two deletion mutations, Delta(61-78) and Delta(121-142),...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi051669r
更新日期:2006-01-31 00:00:00
abstract::Human melanin-concentrating hormone (hMCH) and many of its analogues are potent but nonspecific ligands for human melanin-concentrating hormone receptors 1 and 2 (hMCH-1R and hMCH-2R). To differentiate between the physiological functions of these receptors, selective antagonists are needed. In this study, analogues of...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0200514
更新日期:2002-05-21 00:00:00
abstract::Traditional antibody generation, using either phage display or animal immunization, relies on purified antigens. Many membrane proteins, such as G protein-coupled receptors, solute carriers, or ion channels, are important drug targets but very challenging for the formation of antibodies due to the difficulty of protei...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00730
更新日期:2020-11-10 00:00:00
abstract::The σ1 receptor is an integral membrane protein that shares no homology with other receptor systems, has no unequivocally identified natural ligands, but appears to play critical roles in a wide variety of cell functions. While the number of reports of the possible functions of the σ1 receptor is increasing, almost no...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi401575g
更新日期:2014-05-13 00:00:00
abstract::Equilibrium binding experiments are widely used for the accurate characterization of binding and competitive binding behavior in biological systems. Modern high-throughput discovery efforts in chemical biology rely heavily upon this principle. Here, we derive exact analytical expressions for general competitive bindin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi048233g
更新日期:2004-12-28 00:00:00
abstract::Orthophosphate and phosphate derivatives including pyrophosphate, hexametaphosphate, ATP, ADP, and inositol hexaphosphate enhance the extent of DNA degradation by iron(II) bleomycin. These phosphate-containing compounds increase both the release of free nucleic base and that of base propenals which are DNA cleavage pr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00335a034
更新日期:1985-07-02 00:00:00
abstract::The association-dissociation kinetics of ribosomes from Escherichia coli have been studied under various conditions in a light-scattering stopped-flow apparatus. The dissociation reaction at 2 mg/ml at 25 degrees C, induced by lowering the MgCl2 concentration from 18 to 3 mM, can best be described by three independent...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00671a010
更新日期:1976-12-28 00:00:00
abstract::Specific enzymatic and chemical degradation of neutral lipid and phospholipid fractions from rat liver revealed the presence of novel types of lipid metabolites bearing a short-chain diol backbone. Diol-derived lecithin and cephalin analogs were readily cleaved by phospholipase C (EC 3.1.4.3) from Bacillus cereus, alt...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00675a031
更新日期:1975-02-25 00:00:00
abstract::The Haemophilus influenzae diaminopimelate epimerase was cloned, expressed, purified, and crystallized in the C2221 space group (a = 102.1 A, b = 115.4 A, c = 66.3 A, alpha = beta = gamma = 90 degrees). The three-dimensional structure was solved to 2.7 A using a single Pt derivative and the Se-Met-substituted enzyme t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982138o
更新日期:1998-11-24 00:00:00
abstract::The ligand-binding dynamics and the reaction with O(2) of the fully (five-electron) reduced cytochrome caa(3) from the thermohalophilic bacterium Rhodothermus (R.) marinus were investigated. The enzyme is a proton pump which has all the residues of the proton-transfer pathways found in the mitochondrial-like enzymes c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi010344h
更新日期:2001-09-04 00:00:00
abstract::The discovery of three mutants in the -synuclein (alphaSyn) gene and the identification of alphaSyn as the major component of Lewy body have opened a new field for understanding the pathogenesis of Parkinson's disease (PD). AlphaSyn is a natively unfolded protein with unknown function and unspecified conformational he...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi062068i
更新日期:2007-01-16 00:00:00
abstract::One way to improve the economic feasibility of biomass conversion is to enhance the catalytic efficiency of cellulases through protein engineering. This requires that high-resolution structures of cellulases be available. Here we present the structure of E2cd, the catalytic domain of the thermophilic endocellulase E2 ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00089a006
更新日期:1993-09-28 00:00:00
abstract::The SfiI restriction enzyme binds to DNA as a tetramer holding two usually distant DNA recognition sites together before cleavage of the four DNA strands. To elucidate structural properties of the SfiI-DNA complex, atomic force microscopy (AFM) imaging of the complexes under noncleaving conditions (Ca2+ instead of Mg2...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi051767c
更新日期:2006-01-10 00:00:00
abstract::The empoyment of a novel method of affinity chromatography, which makes use of antibodies that specifically bind DNA/RNA hybrids, has made it possible to enrich for rat rDNA molecules which contain R loops formed with the 18S and 28S rRNAs. An approximately 150-fold enrichment of the ratrRNA coding sequences was obtai...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00580a030
更新日期:1979-06-26 00:00:00
abstract::The active site topography of rabbit CYP4B1 has been studied relative to CYP2B1 and CYP102 using a variety of aromatic probe substrates. Oxidation of the prochiral substrate cumene by CYP4B1, but not CYP2B1 or CYP102, resulted in the formation of the thermodynamically disfavored omega-hydroxy metabolite, 2-phenyl-1-pr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi010395e
更新日期:2001-07-24 00:00:00
abstract::Escherichia coli tyrosine kinase (Etk) regulates the export of pathogenic capsular polysaccharide (CPS) by intermolecularly autophosphorylating its C-terminal tyrosine cluster. The kinase Etk, however, needs to be first activated by the intramolecular phosphorylation of a tyrosine residue, Y574, next to the active sit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi900811p
更新日期:2009-08-25 00:00:00
abstract::A model cosolvent, ethanol, has profound and diversified effects on the amyloidogenic self-assembly of insulin, yielding spectroscopically and morphologically distinguishable forms of beta-aggregates. The alcohol reduces hydrodynamic radii of insulin molecules, decreases enthalpic costs associated with aggregation-pro...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi050281t
更新日期:2005-06-28 00:00:00
abstract::Nucleosome assembly proteins (Naps) influence chromatin dynamics by directly binding to histones. Here we provide a comprehensive structural and biochemical analysis of a Nap protein from Caenorhabditis elegans (CeNap1). CeNap1 naturally lacks the acidic N-terminal tail and has a short C-terminal tail compared to many...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b01033
更新日期:2019-01-15 00:00:00
abstract::We have studied the hydrogen-exchange kinetics of hemoglobin A0 as a function of ligand, CO, saturation. In the noncooperative system, azide binding to methemoglobin, the alterations in exchange kinetics are proportional to the average degree of ligation. However, in the case of CO binding to deoxyhemoglobin the chang...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00297a015
更新日期:1984-01-17 00:00:00
abstract::The repair of the endogenous lesion 8-oxo-7,8-dihydrodeoxyguanosine (8-oxodG) was investigated in the nucleotide excision repair mutant xeroderma pigmentosum D (XPD), using human normal or transformed XPD fibroblasts and the Chinese hamster XPD cell line UV5. In vivo repair of 8-oxodG induced by hydrogen peroxide trea...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992610l
更新日期:2000-08-29 00:00:00
abstract::The mechanisms by which multisubunit histone acetyltransferase (HAT) complexes recognize and perform efficient acetylation on nucleosome substrates are largely unknown. Here, we use a variety of biochemical approaches and compare histone-based substrates of increasing complexity to determine the critical components of...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi602366n
更新日期:2007-02-27 00:00:00