Abstract:
:Chemical modification of mitochondrial F1-ATPase from Schizosaccharomyces pombe by the tryptophan-specific reagent N-bromosuccinimide (NBS) at pH 5.0 in the presence of 20% glycerol produced a characteristic lowering in both enzyme absorbance at 280 nm and intrinsic fluorescence at 332 nm that varied with NBS/F1 molar ratio up to a value of 130. Fluorometric titration of tryptophans and correlation to residual ATPase activity showed that modification of three reactive residues among the seven present on alpha- and epsilon-subunits did not markedly modify the enzyme activity but efficiently released endogenous ATP and abolished the fluorescence quenching related to GDP or ATP binding to the catalytic site. Additional modification of one, less reactive, tryptophan altered both negative cooperativity of ATP hydrolysis and sensitivity to azide inhibition and produced a nearly complete inactivation at high NBS/F1 molar ratio. NBS-induced inactivation of F1 was favored by catalytic-site saturation with GDP or low ATP concentration and on the contrary was prevented by noncatalytic-site saturation with ADP or high ATP concentration. When reactive tryptophans were selectively modified by NBS in the presence of ADP, and subunits were isolated after guanidine hydrochloride dissociation by one-step purification on reversed-phase HPLC, the absorbance of alpha-subunit at 280 nm was decreased, whereas that of epsilon-subunit was unchanged. Cyanogen bromide cleavage of alpha-subunit and fragments separation by reversed-phase HPLC showed that one peptide of 3 kDa apparent molecular mass had decreased absorbance. N-Terminal sequencing allowed its identification to fragment 255-282 that contains tryptophan257.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Divita G,Jault JM,Gautheron DC,Di Pietro Adoi
10.1021/bi00055a004subject
Has Abstractpub_date
1993-02-02 00:00:00pages
1017-24issue
4eissn
0006-2960issn
1520-4995journal_volume
32pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::MutY protein, a DNA glycosylase found in Escherichia coli, recognizes dA:dG, dA:8-oxodG, and dA:dC mismatches in duplex DNA, excising the adenine moiety. We have investigated the mechanism of action of MutY, addressing several points of disagreement raised by previous studies of this enzyme. MutY forms a covalent inte...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi981066y
更新日期:1998-09-08 00:00:00
abstract::A fluorescence method is presented for quantitatively analyzing exocytosis phenomena and monitoring their kinetics. The method is based on the particular properties of a hydrophobic fluorescent probe, 1-[4-(trimethylammonio)phenyl]-6-phenylhexa-1,3,5-triene (TMA-DPH) [Prendergast, F.G., Haugland, R.P., & Callahan, P.J...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00356a045
更新日期:1986-04-22 00:00:00
abstract::Escherichia coli non-heme-binding ferritin A (EcFtnA) is a spherical cagelike protein that is composed of 24 identical subunits. EcFtnA dissociates into 2-mers under acidic conditions and can reassemble into the native structure when the pH is increased. To understand how electrostatic interactions influence the assem...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b01197
更新日期:2016-01-26 00:00:00
abstract::Redox transitions in the Rhodobacter capsulatus cytochrome bc(1) complex were investigated by perfusion-induced attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy combined with synchronous visible spectroscopy, in both the wild type and a cytochrome c(1) point mutant, M183K, in which the mi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049211x
更新日期:2004-07-27 00:00:00
abstract::One of the putative proton-transfer pathways leading from solution toward the binuclear center in many cytochrome c oxidases is the D-pathway, so-called because it starts with a highly conserved aspartate [D(I-132)] residue. Another highly conserved amino acid residue in this pathway, glutamate(I-286), has been indica...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0012641
更新日期:2000-12-26 00:00:00
abstract::We have used nucleobase substitution and kinetic analysis to test the hypothesis that hammerhead catalysis occurs by a general acid-base mechanism, in which nucleobases are directly involved in deprotonation of the attacking 2'-hydroxyl group and protonation of the 5'-oxygen that serves as the leaving group in the cle...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi047941z
更新日期:2005-05-31 00:00:00
abstract::The human translin protein binds a variety of sequences (chromosomal breakpoint consensus sequences, their sequence variants, as well as nonbreakpoint sequences such as simple AT and GC repeats) at nanomolar protein concentration when short single strands ( approximately 20-30mers) are used as DNA targets. The protein...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026378m
更新日期:2002-12-24 00:00:00
abstract::Firefly luciferase is 80-90% inactivated within 3 h upon incubation with the adenine nucleotide analogue p-fluorosulfonylbenzoyl-5'-adenosine (FSBA). Although 4 mol of 14C-FSBA/mol of enzyme is irreversibly bound during inactivation, only 1 mol of 14C-FSBA appears to be specifically directed to an adenine nucleotide b...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00508a031
更新日期:1981-03-03 00:00:00
abstract::The unfolding and refolding properties of human lysozyme and two amyloidogenic variants (Ile56Thr and Asp67His) have been studied by stopped-flow fluorescence and hydrogen exchange pulse labeling coupled with mass spectrometry. The unfolding of each protein in 5.4 M guanidine hydrochloride (GuHCl) is well described as...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi983037t
更新日期:1999-05-18 00:00:00
abstract::Small molecule allostery modifies protein function but is not easily discovered. We introduce mass spectrometry integrated with equilibrium dialysis for the discovery of allostery systematically (MIDAS), a method for identifying physiologically relevant, low-affinity metabolite-protein interactions using unmodified pr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi201313s
更新日期:2012-01-10 00:00:00
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
更新日期:1991-03-19 00:00:00
abstract::We studied the binding of peptides containing five basic residues to membranes containing acidic lipids. The peptides have five arginine or lysine residues and zero, one, or two alanines between the basic groups. The vesicles were formed from mixtures of a zwitterionic lipid, phosphatidylcholine, and an acidic lipid, ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00121a026
更新日期:1992-02-18 00:00:00
abstract::Soluble inorganic pyrophosphatase (PPase) is one of the better understood phosphoryl-transfer enzymes and is distinctive in having four divalent metal ions at the active site. Here we determine pH profiles for wild-type Saccharomyces cerevisiae PPase (Y-PPase) and for 14 of its active site variants and consider the ef...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi971771r
更新日期:1998-02-17 00:00:00
abstract::Centrioles are essential components of the animal centrosome and play crucial roles in the formation of cilia and flagella. They are cylindrical structures composed of nine triplet microtubules organized around a central cartwheel. Recent studies have identified spindle assembly abnormal protein SAS-6 as a critical co...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00978
更新日期:2015-10-20 00:00:00
abstract::The molecular structure of ammonium deoxycytidylyl-(3'-5')-deoxyguanosine, crystallized from aqueous acetone near pH 4, was determined for X-ray diffraction data. The crystals were tetragonal, space group P43212 with a = b = 11.078 (1) A and c = 45.826 (4) A. The structure was solved by tangent expansion of phases bas...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00277a014
更新日期:1983-04-12 00:00:00
abstract::In prokaryotes, a large share of the proteins are secreted from the cell through a process that requires their translocation across the cytoplasmic membrane. This process is mediated by the universally conserved Sec system with homologues in the endoplasmic reticulum and thylakoid membranes of eukaryotes. The Sec syst...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/bi400913x
更新日期:2013-10-01 00:00:00
abstract::Transcripts of Saccharomyces cerevisiae nuclear tRNA genes are normally terminated within a few nucleotides of the tRNA coding region, in contrast to mitochondrially encoded tRNAs, which are contained within polycistronic transcripts and thus require 3'-processing by mitochondrial endonucleases. We show that 3'-proces...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00159a024
更新日期:1992-11-10 00:00:00
abstract::The arrangement of the mRNA on the Escherichia coli ribosome with respect to ribosomal RNA sites has been investigated by photochemical cross-linking experiments. mRNA analogues 51-54 nucleotides in length contained a Shine-Dalgarno sequence, a single codon for tRNA(Gly), and 4-thiouridine (s4U) in the 5' third of the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00176a040
更新日期:1994-03-15 00:00:00
abstract::The respiratory nitric oxide reductase (NOR) from Paracoccus denitrificans catalyzes the two-electron reduction of NO to N(2)O (2NO + 2H(+) + 2e(-) --> N(2)O + H(2)O), which is an obligatory step in the sequential reduction of nitrate to dinitrogen known as denitrification. NOR has four redox-active cofactors, namely,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi050524h
更新日期:2005-08-09 00:00:00
abstract::We investigated the effects of substituting two of the four tryptophans (the "inner pair" Trp(9) and Trp(11) or the "outer pair" Trp(13) and Trp(15)) in gramicidin A (gA) channels. The conformational preferences of the doubly substituted gA analogues were assessed using circular dichroism spectroscopy and size-exclusi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi200136e
更新日期:2011-06-07 00:00:00
abstract::Equilibrium binding isotherms have been measured as a function of temperature for the interaction of 2- and 8-aminoadenosine with poly(U) by continuous, constant-volume ultrafiltration. An analytical method is described for determining the entire isotherm in a single experiment on a preequilibrated ligand--polymer sol...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00530a011
更新日期:1982-01-05 00:00:00
abstract::The extent of rapid (picosecond) backbone motions within the glucocorticoid receptor DNA-binding domain (GR DBD) has been investigated using proton-detected heteronuclear NMR spectroscopy on uniformly 15N-labeled protein fragments containing the GR DBD. Sequence-specific 15N resonance assignments, based on two- and th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00163a007
更新日期:1992-12-08 00:00:00
abstract::The gene for the human substance P receptor (NK-1) was cloned using cDNA probes made by the polymerase chain reaction from primers based on the rat sequence. The gene spans 45-60 kb and is contained in five exons, with introns interrupting at sites homologous to those in the NK-2 receptor gene. Analysis of restriction...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00108a006
更新日期:1991-11-05 00:00:00
abstract::The characteristics of tetrahydrobiopterin (H4biopterin) binding to pteridine-free recombinant macrophage inducible nitric oxide synthase expressed in Escherichia coli were investigated with a special focus given to effects caused by 2,4-diamino-5,6,7, 8-tetrahydro-6-(l-erythro-1,2-dihydroxypropyl)pteridine (4-amino-H...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970144z
更新日期:1997-07-08 00:00:00
abstract::Escherichia coli DNA photolyase contains a stable flavin radical and a second chromophore (SC) of unknown structure. The effects of flash (both conventional and laser) excitation of either the radical alone or both the radical and the second chromophore have been investigated by variation of the excitation wavelengths...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00389a007
更新日期:1987-07-28 00:00:00
abstract::We used CD and UV resonance Raman spectroscopy to study the impact of alcohols on the conformational equilibria and relative Gibbs free energy landscapes along the Ramachandran Psi-coordinate of a mainly poly-Ala peptide, AP with an AAAAA(AAARA)(3)A sequence. 2,2,2-Trifluoroethanol (TFE) most stabilizes the alpha-heli...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi100176a
更新日期:2010-04-20 00:00:00
abstract::The Escherichia coli ribosomal protein S4 is known to repress translation of its own gene and several other ribosomal protein (r-protein) genes in the alpha operon as part of a general mechanism coordinating the levels of rRNA and r-protein synthesis. Using a filter binding assay and RNA transcripts prepared in vitro,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00348a002
更新日期:1985-12-31 00:00:00
abstract::We have validated that ligand peptides designed from antigen peptides could be used for targeting specific major histocompatibility complex class I (MHC-I) molecules on the cell surface. To design the ligand peptides, we used reported antigen peptides for each MHC-I molecule with high binding affinity. From the crysta...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00735
更新日期:2020-12-15 00:00:00
abstract::The tertiary structure of alpha1-acid glycoprotein (AGP) remains unresolved despite its novel function because AGP is a hard target in X-ray and NMR analyses. To elucidate the membrane-induced conformational change of AGP, the vacuum-ultraviolet circular dichroism (VUVCD) spectra of AGP and its constituent sugars were...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901184r
更新日期:2009-09-29 00:00:00
abstract::Our previous studies demonstrated that the first 1000 amino acid residues (the βα1 domain) of human apolipoprotein (apo) B-100, termed apoB:1000, are required for the initiation of lipoprotein assembly and the formation of a monodisperse stable phospholipid (PL)-rich particle. The objectives of this study were (a) to ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.6b01174
更新日期:2017-08-08 00:00:00