Abstract：
:To study the substrate specificity and mechanism of thioltransferase (TTase) catalysis, we have used 14C- and 35S-radiolabeled mixed disulfides of cysteine and glutathione (GSH) with various cysteine-containing proteins. These protein mixed disulfide substrates were incubated with glutathione, glutathione disulfide (GSSG) reductase, and NADPH in the presence or absence of thioltransferase. Glutathione-dependent reduction of protein mixed disulfides was monitored both by release of trichloroacetic acid soluble radiolabel and by formation of GSSG in an NADPH-linked spectrophotometric assay. GSH-dependent dethiolation of [35S]glutathione-papain mixed disulfide (papain-SSG) and the corresponding bovine serum albumin mixed disulfide (BSA-SSG) were catalyzed by thioltransferase (from human red blood cells) as shown by the radiolabel assay, and equivalent rates were measured by the spectrophotometric assay. Dethiolation of [35S]hemoglobin-glutathione mixed disulfide (Hb-SSG) was also catalyzed by TTase. In contrast, TTase did not catalyze GSH-dependent dethiolation of [14C]papain-SScysteine or [14C]BSA-SScysteine as measured by the radiolabel assay. [14C]Hb-SScysteine and Hb-SScysteamine also did not serve as substrates. In separate experiments, TTase from rat liver displayed analogous selectivity. Thus, thioltransferase (glutaredoxin) appears to be specific for glutathione-containing mixed disulfides. Apparent TTase catalysis of GSSG formation from the papain- and BSA-SScysteine mixed disulfides was observed by the spectrophotometric assay, but a lag phase occurred consistent with preenzymatic formation of GSScysteine which could serve as the actual TTase substrate. Two-substrate kinetic studies of TTase with GSH and GSScysteine gave patterns of parallel lines on double-reciprocal plots (1/V vs 1/[S]), consistent with a simple ping-pong mechanism involving a TTase-SSG intermediate.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Gravina SA,Mieyal JJdoi
10.1021/bi00064a021subject
Has Abstractpub_date
1993-04-06 00:00:00pages
3368-76issue
13eissn
0006-2960issn
1520-4995journal_volume
32pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:Carboxyl ester lipase (CEL; EC 3.1.1.13) hydrolyzes cholesteryl esters and retinyl esters in vitro. In vivo, pancreatic CEL is thought to liberate cholesterol and retinol from their esters prior to absorption in the intestine. CEL is also a major lipase in the breast milk of many mammals, including humans and mice, an...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi981679a
更新日期：1999-03-30 00:00:00
abstract：:We have used nucleoside phosphorothioates (NTP alpha S) and a substitution-interference method to identify phosphate oxygens that appear to be important to guanosine cofactor addition in the self-splicing group I intron from Tetrahymena thermophila. For the majority of these phosphate oxygens, however, the effect of N...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00068a001
更新日期：1993-05-04 00:00:00
abstract：:Computer graphics and energy minimization techniques were used to construct a model of DNA containing cis-thymine glycol, an oxidation product of thymine formed in DNA by ionizing radiation. The model simulated an experimental DNA substrate used to study the effects of this lesion on DNA synthesis in vitro. The result...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00391a028
更新日期：1987-08-25 00:00:00
abstract：:During translation, the small subunit of the ribosome rotates with respect to the large subunit primarily between two states as mRNA is being translated into a protein. At the termination of bacterial translation, class I release factors (RFs) bind to a stop codon in the A-site and catalyze the release of the peptide ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00824
更新日期：2018-04-03 00:00:00
abstract：:Infrared (vibrational) circular dichroism (VCD) has been observed for the DNA models d(CG)5, poly(dG-dC).poly(dG-dC), poly(dG).poly(dC), poly(dA-dT).poly(dA-dT), and poly(dA).poly(dT) in the B-conformation in buffered, aqueous solution. The observed results are quantitatively interpreted in terms of the exciton model ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00484a018
更新日期：1990-08-14 00:00:00
abstract：:The photoreceptor protein rhodopsin has been reconstituted with a single phospholipid species, dimyristoylphosphatidylcholine, at a range of different lipid/protein ratios, and the exchange rate at the lipid-protein interface has been determined from the electron spin resonance spectra of spin-labeled phosphatidylchol...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00385a045
更新日期：1987-06-02 00:00:00
abstract：:Thirty single cysteine substitution mutants of T4 lysozyme have been prepared and spin-labeled with a sulfhydryl-specific nitroxide reagent in order to systematically investigate the relationship between nitroxide side-chain mobility and protein structure. The perturbation caused by replacement of a native residue wit...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi960482k
更新日期：1996-06-18 00:00:00
abstract：:Phosphatidic (PtdOH) acid formation is recognized as an important step in numerous signaling pathways in both plants and mammals. To study the role of this lipid in signaling pathways, it is of major interest to be able to increase the amount of this lipid directly. Therefore, "caged" PtdOH was synthesized, which rele...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0351460
更新日期：2004-04-13 00:00:00
abstract：:The dependence of calcineurin on Ca2+ for activity is the result of the concerted action of calmodulin, which increases the turnover rate of the enzyme and modulates its response to Ca2+ transients, and of calcineurin B, which decreases the Km of the enzyme for its substrate. The calmodulin-stimulated protein phosphat...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00188a015
更新日期：1994-06-07 00:00:00
abstract：:The specificity of horse liver alcohol dehydrogenase for cyclohexanol and its 3-methyl derivatives was investigated by stopped-flow and initial velocity kinetic studies. The (1S,3S)-3-methylcyclohexanol was 7 times more reactive (V/Km) than cyclohexanol, whereas the (1R,3R)-3-methylcyclohexanol was at least 1000 times...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00409a060
更新日期：1988-05-03 00:00:00
abstract：:Growth factor receptor-bound protein 14 (Grb14) is involved in growth factor receptor tyrosine kinase signaling. Here we report that light causes a major redistribution of Grb14 among the individual subcellular compartments of the retinal rod photoreceptor. Grb14 is localized predominantly to the inner segment, nuclea...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9000062
更新日期：2009-06-23 00:00:00
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
更新日期：2000-09-05 00:00:00
abstract：:The fluorescence of the base analogue 2-aminopurine (2AP) was used to probe bacteriophage T4 DNA polymerase-induced conformational changes in the template strand produced during the nucleotide incorporation and proofreading reactions. 2AP fluorescence in DNA is quenched by 2AP interactions with neighboring bases, but ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi051462y
更新日期：2005-12-06 00:00:00
abstract：:Recombinant human brain calbindin D(28K) (rHCaBP), human Cu,Zn-superoxide dismutase (HCuZnSOD), rabbit muscle glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and bovine serum albumin (BSA) were found to be S-glutathiolated in decomposed S-nitrosoglutathione (GSNO) solutions. Tryptic or Glu-C digestion and MALDI-TOF ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi035924o
更新日期：2004-04-06 00:00:00
abstract：:Lysine side chains induce conformational changes in plasminogen (Pg) that regulate the process of fibrinolysis or blood clot dissolution. A lysine side-chain mimic, epsilon amino caproic acid (EACA), enhances the activation of Pg by urinary-type and tissue-type Pg activators but inhibits Pg activation induced by strep...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi992028x
更新日期：2000-04-25 00:00:00
abstract：:Light-induced hypsochromic shifts of the Q(y) absorption band of the bacteriochlorophyll dimer (P) from 865 to 850 nm were identified using continuous illumination of dark-adapted reaction centers (RCs) from Rhodobacter capsulatus when dispersed in the most commonly used detergent, the zwitterionic lauryl N-dimethylam...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi200595z
更新日期：2011-06-14 00:00:00
abstract：:Three methods, fluorescence anisotropy of rhodamine-labeled profilin, intrinsic fluorescence and nucleotide exchange, give the same affinity, Kd = 0.1 microM, for Acanthamoeba profilins binding amoeba actin monomers with bound Mg-ATP. Replacement of serine 38 with cysteine created a unique site where labeling with rho...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi980093l
更新日期：1998-08-04 00:00:00
abstract：:The search for a rational method of developing an antimalarial vaccine (malaria caused by Plasmodium falciparum) consists of blocking receptor-ligand interaction. Conserved peptides derived from proteins involved in invasion and having strong red blood cell binding ability have thus been identified; immunization studi...
journal_title：Biochemistry
pub_type： 历史文章,杂志文章
doi：10.1021/bi050214z
更新日期：2005-05-10 00:00:00
abstract：:Kirromycin and related antibiotics inhibit protein synthesis in bacteria by acting on elongation factor Tu (EF-Tu). We have studied the effects of N-methylkirromycin (aurodox) on some molecular properties of this protein. The binding of the antibiotic causes a dramatic variation in the protein fluorescence emission sp...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00443a046
更新日期：1989-08-22 00:00:00
abstract：:Curcumin potentiates the phosphorylation-dependent activity of human cystic fibrosis transmembrane conductance regulator (CFTR) in Fe(3+)-dependent and Fe(3+)-independent manners. Although the Fe(3+)-dependent curcumin potentiation results from removal of endogenous inhibitory Fe(3+) at the interface of the regulatory...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.5b00219
更新日期：2015-05-12 00:00:00
abstract：:In the absence of ligands, dihydrofolate reductase from Escherichia coli exists in at least two interconvertible conformations, only one of which binds NADPH with high affinity. This equilibrium is pH dependent, involving an ionizable group of the enzyme (pK approximately 5.5), and the proportion of the NADPH-binding ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00489a010
更新日期：1990-09-18 00:00:00
abstract：:The effects of detergent [deoxycholate (DOC) and phospholipid [dimyristoylphosphatidylcholine (DMPC)] environments on the rotational dynamics of the single tryptophan residue 26 of bacteriophage M13 coat protein have been investigated by using time-resolved single photon counting measurements of the fluorescence inten...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00441a035
更新日期：1989-07-25 00:00:00
abstract：:N-Linked sugar chains were liberated by hydrazinolysis from porcine zona pellucida glycoproteins obtained from ovarian follicular oocytes. Neutral sugar chains were separated from acidic ones by paper electrophoresis and fractionated with a serial lectin column chromatography and Bio-Gel P-4 column chromatography. The...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00222a012
更新日期：1991-02-26 00:00:00
abstract：:The photocycle of 124 kDa phytochrome A from Avena sativa was studied by Fourier-transform infrared spectroscopy at low temperatures. Difference spectra between the parent state Pr and the intermediates of the Pr-->Pfr pathway, i.e. lumi-R, meta-Ra, and meta-Rc, and between Pfr and the intermediates of the Pfr-->Pr pa...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi960960r
更新日期：1996-08-20 00:00:00
abstract：:The peptides generated from the degradation of the oxidized B chain of bovine insulin by the multiproteinase complex macropain (proteasome) have been analyzed by reverse-phase peptide mapping and identified by N-terminal amino acid sequencing and composition analysis. Six of the 29 peptide bonds in the insulin B chain...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00224a022
更新日期：1991-03-12 00:00:00
abstract：:Self-assembled nanostructures that are sensitive to environmental stimuli are promising nanomaterials for drug delivery. In this class, disulfide-containing redox-sensitive strategies have gained enormous attention because of their wide applicability and simplicity of nanoparticle design. In the context of nucleic aci...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.0c00860
更新日期：2021-01-11 00:00:00
abstract：:Quinone oxidoreductase 2 (QR2) purified from human red blood cells was recently shown to be a potential target of the quinoline antimalarial compounds [Graves et al., (2002) Mol. Pharmacol. 62, 1364]. QR2 catalyzes the two-electron reduction of menadione via the oxidation of N-alkylated or N-ribosylated nicotinamides....
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi035923w
更新日期：2004-04-20 00:00:00
abstract：:Biogenesis of the Gram-negative outer membrane involves the chaperone seventeen kilodalton protein (Skp). A Skp trimer is currently thought to bind its unfolded outer membrane protein (uOMP) substrates. Using sedimentation equilibrium, we discovered that Skp is not an obligate trimer under physiological conditions and...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.5b00806
更新日期：2015-12-08 00:00:00
abstract：:The binding equilibrium of phospholipase A2 (PLA2) to the substrate interface influences many aspects of the overall kinetics of interfacial catalysis by this enzyme. For example, the interpretation of kinetic data on substrate specificity was difficult when there was a significant kinetic contribution from the interf...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00243a037
更新日期：1991-07-23 00:00:00
abstract：:An S3 electron paramagnetic resonance (EPR) signal is observed in a variety of photosystem 2 (PS2) samples in which the oxygen-evolving complex (OEC) has been inhibited. These signals have been proposed to be due to an interaction, S2X+, between the manganese cluster in an oxidation state equivalent to S2 and an organ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00088a032
更新日期：1993-09-21 00:00:00
