scholarly journals TheRhodobacter sphaeroidesmethionine sulfoxide reductase MsrP can reduceR- andS-diastereomers of methionine sulfoxide from a broad-spectrum of protein substrates

2018 ◽  
Author(s):  
Lionel Tarrago ◽  
Sandrine Grosse ◽  
Marina I. Siponen ◽  
David Lemaire ◽  
Béatrice Alonso ◽  
...  
Keyword(s):  
Rhodobacter Sphaeroides ◽  
Broad Spectrum ◽  
Methionine Sulfoxide ◽  
Methionine Sulfoxide Reductase ◽  
Gram Negative Bacteria ◽  
Proteomics Analysis ◽  
E Coli ◽  
Protein Substrates ◽  
Oxidized Proteins ◽  
Periplasmic Proteins

SummaryMethionine (Met) is prone to oxidation and can be converted to Met sulfoxide (MetO), which exists as R- and S-diastereomers. MetO can be reduced back to Met by the ubiquitous methionine sulfoxide reductase (Msr) enzymes. Canonical MsrA and MsrB were shown as absolutely stereospecific for the reduction of S- and R-diastereomer, respectively. Recently, the molybdenum-containing protein MsrP, conserved in all gram-negative bacteria, was shown to be able to reduce MetO of periplasmic proteins without apparent stereospecificity inEscherichia coli.Here, we describe the substrate specificity of theRhodobacter sphaeroidesMsrP. Proteomics analysis coupled to enzymology approaches indicate that it reduces a broad spectrum of periplasmic oxidized proteins. Moreover, using model proteins, we demonstrated that RsMsrP preferentially reduces unfolded oxidized proteins and we confirmed that this enzyme, like itsE. colihomolog, can reduce bothR-andS-diastereomers of MetO with similar efficiency.

scholarly journals Rhodobacter sphaeroides methionine sulfoxide reductase P reduces R- and S-diastereomers of methionine sulfoxide from a broad-spectrum of protein substrates

2018 ◽  
Vol 475 (23) ◽  
pp. 3779-3795 ◽  
Author(s):  
Lionel Tarrago ◽  
Sandrine Grosse ◽  
Marina I. Siponen ◽  
David Lemaire ◽  
Béatrice Alonso ◽  
...  
Keyword(s):  
Rhodobacter Sphaeroides ◽  
Broad Spectrum ◽  
Reducing Power ◽  
Methionine Sulfoxide ◽  
Biochemical Properties ◽  
Amino Acid Sequences ◽  
Methionine Sulfoxide Reductase ◽  
Enzymatic System ◽  
Protein Substrates ◽  
Electron Transport Chains

Methionine (Met) is prone to oxidation and can be converted to Met sulfoxide (MetO), which exists as R- and S-diastereomers. MetO can be reduced back to Met by the ubiquitous methionine sulfoxide reductase (Msr) enzymes. Canonical MsrA and MsrB were shown to be absolutely stereospecific for the reduction of S-diastereomer and R-diastereomer, respectively. Recently, a new enzymatic system, MsrQ/MsrP which is conserved in all gram-negative bacteria, was identified as a key actor for the reduction of oxidized periplasmic proteins. The haem-binding membrane protein MsrQ transmits reducing power from the electron transport chains to the molybdoenzyme MsrP, which acts as a protein-MetO reductase. The MsrQ/MsrP function was well established genetically, but the identity and biochemical properties of MsrP substrates remain unknown. In this work, using the purified MsrP enzyme from the photosynthetic bacteria Rhodobacter sphaeroides as a model, we show that it can reduce a broad spectrum of protein substrates. The most efficiently reduced MetO is found in clusters, in amino acid sequences devoid of threonine and proline on the C-terminal side. Moreover, R. sphaeroides MsrP lacks stereospecificity as it can reduce both R- and S-diastereomers of MetO, similarly to its Escherichia coli homolog, and preferentially acts on unfolded oxidized proteins. Overall, these results provide important insights into the function of a bacterial envelop protecting system, which should help understand how bacteria cope in harmful environments.

scholarly journals HypVW is an HOCl-Sensing Two Component System in Escherichia coli

2021 ◽  
Author(s):  
Sara El Hajj ◽  
Camille Henry ◽  
Alexandra Vergnes ◽  
Laurent Loiseau ◽  
Brasseur Gael ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Hypochlorous Acid ◽  
Methionine Sulfoxide ◽  
Methionine Sulfoxide Reductase ◽  
Bacterial Cells ◽  
E Coli ◽  
Two Component ◽  
Redox Switch ◽  
Two Component Systems ◽  
Methionine Residues

Two component systems (TCS) are signalling pathways that allow bacterial cells to sense, respond and adapt to fluctuating environments. Among the classical TCS of Escherichia coli, YedVW has been recently showed to be involved in the regulation of msrPQ, encoding for the periplasmic methionine sulfoxide reductase system. In this study, we demonstrate that hypochlorous acid (HOCl) induces the expression of msrPQ in a YedVW dependant manner, whereas H2O2, NO and paraquat (a superoxide generator) do not. Therefore, YedV appears to be an HOCl-sensing histidine kinase. Based on this finding, we proposed to rename this system HypVW.  Moreover, using a directed mutagenesis approach, we show that Met residues located in the periplasmic loop of HypV (formerly YedV) are important for its activity. Given that HOCl oxidizes preferentially Met residues, we bring evidences that HypV could be activated via the reversible oxidation of its methionine residues, thus conferring to MsrPQ a role in switching HypVW off. Based on these results, we propose that the activation of HypV by HOCl could occur through a Met redox switch. HypVW appears to be the first characterized TCS able to detect HOCl in E. coli. This study represents an important step in understanding the mechanisms of reactive chlorine species resistance in prokaryotes.

scholarly journals Competitive cobalt for zinc substitution in mammalian methionine sulfoxide reductase B1 overexpressed in E. coli: structural and functional insight

2013 ◽  
Vol 19 (1) ◽  
pp. 85-95 ◽  
Author(s):  
Elena Shumilina ◽  
Olena Dobrovolska ◽  
Rebecca Del Conte ◽  
Henrik Waldal Holen ◽  
Alexander Dikiy
Keyword(s):  
Methionine Sulfoxide ◽  
Methionine Sulfoxide Reductase ◽  
E Coli

scholarly journals Methionine Sulfoxide Reduction and Assimilation in Escherichia coli: New Role for the Biotin Sulfoxide Reductase BisC

2005 ◽  
Vol 187 (1) ◽  
pp. 231-237 ◽  
Author(s):  
Benjamin Ezraty ◽  
Julia Bos ◽  
Frédéric Barras ◽  
Laurent Aussel
Keyword(s):  
Escherichia Coli ◽  
Essential Role ◽  
Reductase Activity ◽  
Methionine Sulfoxide ◽  
Mass Spectrometric ◽  
Methionine Sulfoxide Reductase ◽  
Racemic Mixture ◽  
E Coli ◽  
Methionine Sulfoxide Reductases ◽  
Biotin Sulfoxide Reductase

ABSTRACT Methionine ranks among the amino acids most sensitive to oxidation, which converts it to a racemic mixture of methionine-S-sulfoxide (Met-S-SO) and methionine-R-sulfoxide (Met-R-SO). The methionine sulfoxide reductases MsrA and MsrB reduce free and protein-bound MetSO, MsrA being specific for Met-S-SO and MsrB for Met-R-SO. In the present study, we report that an Escherichia coli metB1 auxotroph lacking both msrA and msrB is still able to use either of the two MetSO enantiomers. This indicates that additional methionine sulfoxide reductase activities occur in E. coli. BisC, a poorly characterized biotin sulfoxide reductase, was identified as one of these new methionine sulfoxide reductases. BisC was purified and found to exhibit reductase activity with free Met-S-SO but not with free Met-R-SO as a substrate. Moreover, a metB1 msrA msrB bisC strain of E. coli was unable to use Met-S-SO for growth, but it retained the ability to use Met-R-SO. Mass spectrometric analyses indicated that BisC is unable to reduce protein-bound Met-S-SO. Hence, this study shows that BisC has an essential role in assimilation of oxidized methionines. Moreover, this work provides the first example of an enzyme that reduces free MetSO while having no activity on peptide-bound MetSO residues.

scholarly journals YbfA Regulates the Sensitivity of Escherichia coli K12 to Plantaricin BM-1 via the BasS/BasR Two-Component Regulatory System

2021 ◽  
Vol 12 ◽  
Author(s):  
Xinyue Chen ◽  
Yifei Liu ◽  
Junhua Jin ◽  
Hui Liu ◽  
Yanling Hao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Membrane ◽  
Metabolic Pathways ◽  
Gram Negative Bacteria ◽  
Two Component System ◽  
Proteomics Analysis ◽  
Component System ◽  
E Coli ◽  
Two Component ◽  
And Function

Plantaricin BM-1, a class IIa bacteriocin produced by Lactobacillus plantarum BM-1, shows obvious antibacterial activity against Escherichia coli. However, the mechanism underlying the action of class IIa bacteriocins against gram-negative bacteria remains to be explored. The purpose of this study was to investigate the role of YbfA, a DUF2517 domain-containing protein, in the response of Escherichia coli K12 to plantaricin BM-1. The growth curve experiment and MIC experiment showed that the sensitivity of E. coli to plantaricin BM-1 was decreased by a ybfA null mutation. Electron microscopy showed that the ybfA null mutation reduced the surface rupture and contraction caused by plantaricin BM-1, and mitigated the effect of plantaricin BM-1 on the morphology of the E. coli cell membrane. Proteomics analysis showed that 323 proteins were differentially expressed in E. coli lacking the ybfA gene (P < 0.05); 118 proteins were downregulated, and 205 proteins were upregulated. The metabolic pathways containing the upregulated proteins mainly included outer membrane proteins, integral components of the plasma membrane, regulation of cell motility, and regulation of locomotion. The metabolic pathways involving the downregulated proteins mainly included outer membrane protein glycine betaine transport, amino-acid betaine transport, and transmembrane signaling receptor activity. The results of the proteomics analysis showed that the protein expression of the BasS/BasR two-component system was significantly increased (P < 0.05). Moreover, the expression levels of downstream proteins regulated by this two-component system were also significantly increased, including DgkA, FliC, and MlaE, which are involved in cell membrane structure and function, and RT-qPCR also confirmed this result. The growth curve showed that the sensitivity of E. coli to plantaricin BM-1 was significantly increased due to deletion of the BasS/BasR two-component system. Thus, deletion of ybfA in E. coli can increase the expression of the BasS/BasR two-component system and positively regulate the structure and function of the cell membrane to reduce the sensitivity to plantaricin BM-1. This will help to explore the mechanism of action of class IIa bacteriocins against gram-negative bacteria.

Mitochondrial targeting of the human peptide methionine sulfoxide reductase (MSRA), an enzyme‐involved in the repair of oxidized proteins

2002 ◽  
Vol 16 (8) ◽  
pp. 911-913 ◽  
Author(s):  
Alfred Hansel ◽  
Lioba Kuschel ◽  
Solveig Hehl ◽  
Cornelius Lemke ◽  
Hans‐Jürgen Agricola ◽  
...  
Keyword(s):  
Methionine Sulfoxide ◽  
Methionine Sulfoxide Reductase ◽  
Mitochondrial Targeting ◽  
Oxidized Proteins ◽  
Peptide Methionine Sulfoxide Reductase

scholarly journals BioSAXS measurements reveal that two antimicrobial peptides induce similar molecular changes in Gram-negative and Gram-positive bacteria

2019 ◽  
Author(s):  
A.R. von Gundlach ◽  
M. Ashby ◽  
J. Gani ◽  
P. M. Lopez-Perez ◽  
A. Cookson ◽  
...  
Keyword(s):  
Broad Spectrum ◽  
Mode Of Action ◽  
Bacterial Load ◽  
Ultrastructural Changes ◽  
Resistant Bacteria ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
E Coli ◽  
Gram Positive Bacteria

AbstractTwo highly active short broad-spectrum AMPs (14D and 69D) with unknown mode of action have been investigated in regards to their effect against the Gram-negative bacteriaE. coliand the Gram-positive bacteria methicillin-resistantStaphylococcus aureus(MRSA). Minimal inhibitory concentration (MIC) measurements using a cell density of 108cfu/ml resulted in values between 16 and 32 μg/ml. Time kill experiments using 108cfu/ml revealed complete killing, except for 69D in combination with MRSA, where bacterial load was reduced a million times. Small angle X-ray scattering of biological samples (BioSAXS) at 108cfu/ml was applied to investigate the ultrastructural changes inE. coliand MRSA in response to these two broad-spectrum AMPs. In addition, electron microscopy (EM) was performed to visualize the treated and non-treated bacteria. As expected, the scattering curves generated using BioSAXS show the ultrastructure of the Gram-positive and Gram-negative bacteria to be very different (BioSAXS is not susceptible to the outer shape). After treatment with either peptide, the scattering curves ofE. coliand MRSA cells are much more alike. This data in conjunction with the EM indicates that ribosomes might be effected by the treatment as well as changes in the nucleoid occurs. Whereas in EM it is notoriously difficult to observe changes for spherical Gram-positives, the BioSAXS results are superior and reveal strongly similar effects for both peptides induced in Gram-positive as well as Gram-negative bacteria. Given the high-throughput possibility and robust statistics BioSAXS can support and speed up mode of action research in AMPs and other antimicrobial compounds, making a contribution towards the development of urgently needed drugs against resistant bacteria.

Sign in / Sign up

Export Citation Format

Share Document