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1.
Methionine (Met) is an essential sulfur‐containing amino acid, sensitive to oxidation. The oxidation of Met can occur by numerous pathways, including enzymatic modifications and oxidative stress, being able to cause relevant alterations in protein functionality. Under UV radiation, Met may be oxidized by direct absorption (below 250 nm) or by photosensitized reactions. Herein, kinetics of the reaction and identification of products during photosensitized oxidation were analyzed to elucidate the mechanism for the degradation of Met under UV‐A irradiation using pterins, pterin (Ptr) and 6‐methylpterin (Mep), as sensitizers. The process begins with an electron transfer from Met to the triplet‐excited state of the photosensitizer (Ptr or Mep), to yield the corresponding pair of radicals, Met radical cation (Met?+) and the radical anion of the sensitizer (Sens??). In air‐equilibrated solutions, Met?+ incorporates one or two atoms of oxygen to yield methionine sulfoxide (MetO) and methionine sulfone (MetO2), whereas Sens?? reacts with O2 to recover the photosensitizer and generate superoxide anion (O2??). In anaerobic conditions, further free‐radical reactions lead to the formation of the corresponding dihydropterin derivatives (H2Ptr or H2Mep).  相似文献   

2.
Flax cyclic peptides (orbitides, linusorbs (LOs)) [1–8‐NαC],[1‐MetO2]‐linusorb B1 ([MetO2]‐LO1) and [1–9‐NαC],[1‐MetO2]‐linusorb B2 ([MetO2]‐LO2) are biologically active. These LOs lack active nuclei commonly used in peptide modification. We have developed reactions to activate methionine methyl sulphide to produce stable derivatives. In these reactions, LOs are converted to sulfonium intermediates and subsequently to derivatives containing active nuclei while preserving their fundamental structures. The reaction conditions preserved cyclic peptide fundamental structure and organic solvent solubility. [Met]‐LO1 and [Met]‐LO2 analogues containing activated groups (?CN, ?COOEt, and ?NH2) in the form of methionine, methionine (S)‐oxide, and methionine (S,S)‐dioxide amino acids were synthesized and characterized by LCMS and 1D and 2D NMR spectroscopy. Coumarin orbitide complexes produced in this manner bind Eu3+ yielding FRET compounds that absorb energy through coumarin antennae and emit photons at lanthanide wavelengths.  相似文献   

3.
The [Fc? bis{ZnII(TACN)(Py)}] complex, comprising two ZnII(TACN) ligands (Fc=ferrocene; Py=pyrene; TACN=1,4,7‐triazacyclononane) bearing fluorescent pyrene chromophores linked by an electrochemically active ferrocene molecule has been synthesised in high yield through a multistep procedure. In the absence of the polyphosphate guest molecules, very weak excimer emission was observed, indicating that the two pyrene‐bearing ZnII(TACN) units are arranged in a trans‐like configuration with respect to the ferrocene bridging unit. Binding of a variety of polyphosphate anionic guests (PPi and nucleotides di‐ and triphosphate) promotes the interaction between pyrene units and results in an enhancement in excimer emission. Investigations of phosphate binding by 31P NMR spectroscopy, fluorescence and electrochemical techniques confirmed a 1:1 stoichiometry for the binding of PPi and nucleotide polyphosphate anions to the bis(ZnII(TACN)) moiety of [Fc? bis{ZnII(TACN)(Py)}] and indicated that binding induces a trans to cis configuration rearrangement of the bis(ZnII(TACN)) complexes that is responsible for the enhancement of the pyrene excimer emission. Pyrophosphate was concluded to have the strongest affinity to [Fc? bis{ZnII(TACN)(Py)}] among the anions tested based on a six‐fold fluorescence enhancement and 0.1 V negative shift in the potential of the ferrocene/ferrocenium couple. The binding constant for a variety of polyphosphate anions was determined from the change in the intensity of pyrene excimer emission with polyphosphate concentration, measured at 475 nm in CH3CN/Tris‐HCl (1:9) buffer solution (10.0 mM , pH 7.4). These measurements confirmed that pyrophosphate binds more strongly (Kb=(4.45±0.41)×106 M ?1) than the other nucleotide di‐ and triphosphates (Kb=1–50×105 M ?1) tested.  相似文献   

4.
S ‐Adenosylmethionine (SAM) is one of the most common co‐substrates in enzyme‐catalyzed methylation reactions. Most SAM‐dependent reactions proceed through an SN2 mechanism, whereas a subset of them involves radical intermediates for methylating non‐nucleophilic substrates. Herein, we report the characterization and mechanistic investigation of NosN, a class C radical SAM methyltransferase involved in the biosynthesis of the thiopeptide antibiotic nosiheptide. We show that, in contrast to all known SAM‐dependent methyltransferases, NosN does not produce S ‐adenosylhomocysteine (SAH) as a co‐product. Instead, NosN converts SAM into 5′‐methylthioadenosine as a direct methyl donor, employing a radical‐based mechanism for methylation and releasing 5′‐thioadenosine as a co‐product. A series of biochemical and computational studies allowed us to propose a comprehensive mechanism for NosN catalysis, which represents a new paradigm for enzyme‐catalyzed methylation reactions.  相似文献   

5.
A rapid, selective and sensitive hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass spectrometry (MS/MS) method was developed to determine 1‐13C‐l ‐methionine in rat serum. Proteins in serum were precipitated using acetonitrile and the supernatant was separated after centrifugation. 1‐13C‐l ‐phenylalnine was used as the internal standard. HILIC–tandem mass spectrometry analysis was performed on a hydrophilic interaction silica column (TSK‐GEL AMIDE‐80) using a linear gradient elution system, acetonitrile−5 mm ammonium acetate containing 0.1% formic acid and multiple reaction monitoring mode for 1‐13C‐l ‐methionine and 1‐13C‐l ‐phenylalnine. The assay was validated with a linear range between 10 and 150 ng mL−1 (r ≥ 0.99) and a lower limit of quantification of 10 ng mL−1, calculated with weighted (1/x2) least squares linear regression. The RSD of intra‐day precision was smaller than 3.6% and the inter‐day RSD less than 6.5%, while the average recovery was 100.48% with an RSD of accuracy within 2.9%, determined from quality control samples. The HILIC‐MS/MS method was fully validated and successfully applied to the in vivo pharmacokinetic study of stable‐isotope 1‐13C‐l ‐methionine in rats. Copyright © 2011 John Wiley & Sons, Ltd.  相似文献   

6.
The C‐terminal octapeptide of cholecystokinin (CCK8) includes some easily oxidizable amino acids. The oxidation of CCK8 by reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) and hydroxyl radicals (OH?) was investigated using reversed‐phase high performance liquid chromatography (RP‐HPLC) and subsequent electrospray ionization mass spectrometry. The mechanism of oxidation of CCK8 in the H2O2 system differed from that of CCK8 in the Fenton system, in which OH? are produced. In the H2O2 system, 28Met and 31Met were oxidized to methionine sulfoxide, and no further oxidation or degradation/hydrolysis occurred. On the other hand, in the Fenton system, 28Met and 31Met residues were oxidized to methionine sulfone via the formation of methionine sulfoxide. In addition, the oxidized product was observed at the Trp residue but not at the Tyr residue, and small peptide fragments from CCK8 were observed in the Fenton system. From these results, it was concluded that 28Met and 31Met residues of CCK8 are susceptible to oxidation by ROS. Copyright © 2009 John Wiley & Sons, Ltd.  相似文献   

7.
In phosphate buffer media (pH 5.8–8.0), bisperoxo(1,10‐phenanthroline)oxovanadate(V) ( 1 ) oxidizes l ‐methionine to methionine sulfoxide. The stoichiometry of the reaction is 1:1. The reaction occurs in two subsequent first‐order steps. In the first step, one of the peroxo ligands of 1 gets substituted by l ‐methionine. The observed first‐order rate constants for both steps increase linearly with increasing [H+] as well as with increasing [l ‐methionine]. The EPR spectra prove that the reaction involves a cysteinyl radical‐type intermediate and that VV gets reduced to a VIV species.  相似文献   

8.
The usefulness of a C60‐fullerene modified gold (Au) electrode in mediating the oxidation of methionine in the presence of potassium ions electrolyte has been demonstrated. During cyclic voltammetry, an oxidation peak of methionine appearing at +1.0 V vs. Ag/AgCl was observed. The oxidation current of methionine is enhanced by about 2 times using a C60 modified gold electrode. The current enhancement is significantly dependent on pH, temperature and C60 dosage. Calibration plot reveals linearity of up to 0.1 mM with a current sensitivity of close to 50 mA L mol?1 and detection limit of 8.2×10?6 M. The variation of scan rate study shows that the system undergoes diffusion‐controlled process. Diffusion coefficient and rate constant of methionine were determined using hydrodynamic method (rotating disk electrode) with values of 1.11×10?5 cm2 s?1 and 0.0026 cm s?1 respectively for unmodified electrode while the values of diffusion coefficient and rate constant of methionine using C60 modified Au electrode are 5.7×10?6 cm2 s?1 and 0.0021 cm s?1 respectively.  相似文献   

9.
Single crystals of mercury(II) polyphosphate, Hg(PO3)2, were prepared from HgO in an acidic polyphosphate melt. The structure is isotypic with α‐Cd(PO3)2 and comprises infinite polyphosphate chains with a period of four phosphate units. Chains of the form 1[PO3?] are linked by Hg2+ to form a three‐dimensional network. The Hg atom is located at the centre of a distorted octahedron of O atoms with distances 2.173 (5) < (Hg—O)mean < 2.503 (6) Å. The [HgO6] polyhedra form zigzag‐like chains of the form 1[HgO2O4/2] parallel to the c axis.  相似文献   

10.
The synthesis in one‐pot reactions and structural characterization of six new tri‐n‐butyltin(IV) derivatives of Schiff bases are reported. The compounds are derived from a condensation reaction between l ‐alanine, l ‐valine, l ‐isoleucine, l ‐methionine, l ‐phenylalanine or l ‐tryptophan and 3,5‐di‐tert‐butyl‐2‐hydroxybenzaldehyde. Characterization was completed using elemental analysis, infrared spectroscopy, mass spectrometry, one‐ and two‐dimensional solution NMR (1H, 13C and 119Sn) as well as solid‐state 119Sn NMR. In addition, the crystal structures of three of the compounds were confirmed using single‐crystal X‐ray diffraction. Although five‐coordinated and polymeric in the solid state, the tin compounds are four‐coordinated and monomeric in solution. The coordination environment around the triorganotin units comprises three carbon atoms and two oxygen atoms from two ligands in a trigonal bipyramidal geometry. The anti‐proliferative effect of these compounds on the cervical carcinoma cell lines HeLa, CaSki and ViBo was screened in vitro, the compounds showing cytotoxic activity against all three strains and null or low cytotoxic activity (necrotic) as well. Copyright © 2016 John Wiley & Sons, Ltd.  相似文献   

11.
Methylation and demethylation of DNA, RNA and proteins has emerged as a major regulatory mechanism. Studying the function of these modifications would benefit from tools for their site‐specific inhibition and timed removal. S‐Adenosyl‐L‐methionine (AdoMet) analogs in combination with methyltransferases (MTases) have proven useful to map or block and release MTase target sites, however their enzymatic generation has been limited to aliphatic groups at the sulfur atom. We engineered a SAM synthetase from Cryptosporidium hominis (PC‐ChMAT) for efficient generation of AdoMet analogs with photocaging groups that are not accepted by any WT MAT reported to date. The crystal structure of PC‐ChMAT at 1.87 Å revealed how the photocaged AdoMet analog is accommodated and guided engineering of a thermostable MAT from Methanocaldococcus jannaschii. PC‐MATs were compatible with DNA‐ and RNA‐MTases, enabling sequence‐specific modification (“writing”) of plasmid DNA and light‐triggered removal (“erasing”).  相似文献   

12.
ZnS/ZnAl2S4 nanocomposite and 2‐chlorobenzoyl ferrocene, were synthesized and used to construct a modified carbon paste electrode. The electrooxidation of methionine at the surface of the modified electrode was studied. Under the optimized conditions, the square wave voltammetric (SWV) peak current of methionine increased linearly with methionine concentration in the range of 5.0×10?8 to 8.0×10?4 M and detection limit of 10.0 nM was obtained for methionine. The prepared modified electrode exhibits a very good resolution between the voltammetric peaks of methionine and tryptophan which makes it suitable for the detection of methionine in the presence of tryptophan in real samples.  相似文献   

13.
S‐adenosyl‐l ‐methionine (SAM)‐dependent methyltransferases (MTs) catalyse the methylation of a vast array of small metabolites and biomacromolecules. Recently, rare carboxymethylation pathways have been discovered, including carboxymethyltransferase enzymes that utilise a carboxy‐SAM (cxSAM) cofactor generated from SAM by a cxSAM synthase (CmoA). We show how MT enzymes can utilise cxSAM to catalyse carboxymethylation of tetrahydroisoquinoline (THIQ) and catechol substrates. Site‐directed mutagenesis was used to create orthogonal MTs possessing improved catalytic activity and selectivity for cxSAM, with subsequent coupling to CmoA resulting in more efficient and selective carboxymethylation. An enzymatic approach was also developed to generate a previously undescribed co‐factor, carboxy‐S‐adenosyl‐l ‐ethionine (cxSAE), thereby enabling the stereoselective transfer of a chiral 1‐carboxyethyl group to the substrate.  相似文献   

14.
Methionine is an essential amino acid and is the primary source of sulfur for humans. Using the double spike (33S‐36S) multi‐collector thermal ionization mass spectrometry (MC‐TIMS) technique, three sample bottles of a methionine material obtained from the Institute for Reference Materials and Measurements have been measured for δ34S and sulfur concentration. The mean δ34S value, relative to Vienna Canyon Diablo Troilite (VCDT), determined was 10.34 ± 0.11‰ (n = 9) with the uncertainty reported as expanded uncertainties (U). These δ34S measurements include a correction for blank which has been previously ignored in studies of sulfur isotopic composition. The sulfur concentrations for the three bottles range from 56 to 88 µg/g. The isotope composition and concentration results demonstrate the high accuracy and precision of the DS‐MC‐TIMS technique for measuring sulfur in methionine. Published in 2010 by John Wiley & Sons, Ltd.  相似文献   

15.
The early events in the triplet 4-carboxybenzophenone (CB)-induced oxidation of N-acetyl-methionine methyl ester (N-Ac-Met-OCH3) are investigated in aqueous solution. Upon electron transfer from the methionine residue of N-Ac-Met-OCH3 to 3CB*, the resulting sulfur radical cation undergoes further reactions: (1) back electron transfer, (2) escape of the radical ions from the solvent cage, or (3) proton transfer and escape of the radicals. The yields and paths of these reactions are shown to depend strongly on the pH of the solution, and, similar to the previously reported results for dipeptides (Met-Gly and Gly-Met), on the structural nature of the methionine substituents. In the experiments performed in this work, low quencher concentrations were used to avoid formation of intermolecular transients (e.g., dimeric sulfur-centered radical cation (S∴S)+). Under these experimental conditions, the one-electron oxidized sulfur does not seem to become stabilized in an (S∴N)+ three-electron bonded intramolecular complex. The proposed mechanism is further supported by the stable products analysis. A detailed mechanism involving characterization of the transients is discussed and compared to that of methionine and methionine-containing dipeptides (Met-Gly and Gly-Met). Moreover, a newly installed transient absorption laser system is described in details.  相似文献   

16.
The oxidation of methionine (Met) plays an important role during biological conditions of oxidative stress as well as for protein stability. Ruthenium(III)–polypyridyl complexes, [Ru(NN)3]3+, generated from the photochemical oxidation of the corresponding Ru(II) complexes with molecular oxygen, undergo a facile electron transfer reaction with Met to form methionine sulfoxide (MetO) as the final product. Interaction of [Ru(NN)3]3+ with methionine leads to the formation of >S+● and (>S∴S<)+ species as intermediates during the course of the reaction. The interesting spectral, kinetic, and mechanistic study of the electron transfer reaction of four substituted methionines with six [Ru(NN)3]3+ ions carried out in aqueous CH3CN (1:1, v/v) by a spectrophotometric technique shows that the reaction rate is susceptible to the nature of the ligand in [Ru(NN)3]3+ and the structure of methionine. The rate constants calculated by the application of Marcus semiclassical theory to these redox reactions are in close agreement with the experimental values.  相似文献   

17.
Crystallographic analysis of a solid solution of two diastereoisomers, i.e. ({(1S,R)‐1‐carboxy‐3‐[(R,S)‐methylsulfinyl]propyl}aminocarbonyl)methanaminium tetrachloridoaurate(III) and ({(1S,R)‐1‐carboxy‐3‐[(S,R)‐methylsulfinyl]propyl}aminocarbonyl)methanaminium tetrachloridoaurate(III), (C7H15N2O4S)[AuCl4], has shown that in the presence of gold(III), the methionine part of the Gly‐d ,l ‐Met dipeptide is oxidized to sulfoxide, and no coordination to the AuIII cation through the S atom of the sulfoxide is observed. In view of our observation, literature reports that methionine acts as an N,S‐bidentate donor ligand forming stable gold(III) complexes require verification. Moreover, it has been demonstrated that crystallization of the oxidation product leads to a substantial 77:23 excess of both S‐methionine/R‐sulfoxide and R‐methionine/S‐sulfoxide over S‐methionine/S‐sulfoxide and R‐methionine/R‐sulfoxide. The presence of two different diastereoisomers at the same crystallographic site is a source of static disorder at this site.  相似文献   

18.
近年来,水溶性聚磷酸铵在液体肥料和复合肥料的领域受到了广泛的关注,并在发达国家中得到了大面积的推广及应用。在pH值为5.5~8.0、温度为278.15 K~323.15 K的条件下,本文采用滴定法研究Ca2+-Mg2+-Zn2+体系在聚磷酸铵溶液中的螯合规律。实验结果表明:相同质量分数的聚磷酸铵溶液对金属离子的螯合量会随着体系中Ca2+、Mg2+、Zn2+的摩尔浓度的变化而变化;随着温度的升高而逐渐降低;随着pH的增加而逐渐增加;随着聚合度的升高而逐渐增加。采用傅里叶红外光谱对聚磷酸铵和A1B3C3体系的螯合物进行表征。  相似文献   

19.
S ‐Adenosylmethionine‐dependent methyltransferases (MTs) play a decisive role in the biosynthesis of natural products and in epigenetic processes. MTs catalyze the methylation of heteroatoms and even of carbon atoms, which, in many cases, is a challenging reaction in conventional synthesis. However, C‐MTs are often highly substrate‐specific. Herein, we show that SgvM from Streptomyces griseoviridis features an extended substrate scope with respect to the nucleophile as well as the electrophile. Aside from its physiological substrate 4‐methyl‐2‐oxovalerate, SgvM catalyzes the (di)methylation of pyruvate, 2‐oxobutyrate, 2‐oxovalerate, and phenylpyruvate at the β‐carbon atom. Chiral‐phase HPLC analysis revealed that the methylation of 2‐oxovalerate occurs with R selectivity while the ethylation of 2‐oxobutyrate with S ‐adenosylethionine results in the S enantiomer of 3‐methyl‐2‐oxovalerate. Thus SgvM could be a valuable tool for asymmetric biocatalytic C‐alkylation reactions.  相似文献   

20.
Methyltransferases have proven useful to install functional groups site‐specifically in different classes of biomolecules when analogues of their cosubstrate S‐adenosyl‐l ‐methionine (AdoMet) are available. Methyltransferases have been used to address different classes of RNA molecules selectively and site‐specifically, which is indispensable for biophysical and mechanistic studies as well as labeling in the complex cellular environment. However, the AdoMet analogues are not cell‐permeable, thus preventing implementation of this strategy in cells. We present a two‐step enzymatic cascade for site‐specific mRNA modification starting from stable methionine analogues. Our approach combines the enzymatic synthesis of AdoMet with modification of the 5′ cap by a specific RNA methyltransferase in one pot. We demonstrate that a substrate panel including alkene, alkyne, and azido functionalities can be used and further derivatized in different types of click reactions.  相似文献   

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