Overexpression of SRO_3163, a homolog of Streptomyces antibiotic regulatory protein, induces the production of novel cyclohexene-containing enamide in Streptomyces rochei

2021 ◽  
Author(s):  
Yuya Misaki ◽  
Yosi Nindita ◽  
Kota Fujita ◽  
Amirudin Akhmad Fauzi ◽  
Kenji Arakawa
Keyword(s):  
Secondary Metabolites ◽  
Chemical Synthesis ◽  
Active Compound ◽  
Parent Strain ◽  
Regulatory Protein ◽  
Regulatory Proteins ◽  
Molecular Formula ◽  
Transcriptional Activators ◽  
Nmr Analysis ◽  
Streptomyces Species

Abstract Streptomyces antibiotic regulatory proteins (SARPs) are well characterized as transcriptional activators for secondary metabolites in Streptomyces species. Streptomyces rochei 7434AN4 harbors 15 SARP genes, among which 3 were located on a giant linear plasmid pSLA2-L and others were on the chromosome. Some SARP genes were cloned into an integrative thiostrepton-inducible vector pIJ8600, and their recombinants were cultivated. The recombinant of SARP gene, SRO_3163, accumulated a UV-active compound YM3163-A, which was not detected in the parent strain and other SARP recombinants. Its molecular formula was established to be C8H11NO. Extensive NMR analysis revealed that YM3163-A is a novel enamide, 2-(cyclohex-2-en-1-ylidene)acetamide, and its structure was confirmed by chemical synthesis including Horner-Wadsworth-Emmons reaction and ammonolysis.

scholarly journals Isolation, Characterization and Chemical Synthesis of Large Spectrum Antimicrobial Cyclic Dipeptide (l-leu-l-pro) from Streptomyces misionensis V16R3Y1 Bacteria Extracts. A Novel 1H NMR Metabolomic Approach

Antibiotics ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 270
Author(s):  
Ilhem Saadouli ◽  
Imène Zendah El Euch ◽  
Emna Trabelsi ◽  
Amor Mosbah ◽  
Alaeddine Redissi ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Chemical Synthesis ◽  
1H Nmr ◽  
Pathogenic Bacteria ◽  
Antimicrobial Activities ◽  
Biologically Active ◽  
Penicillium Expansum ◽  
Cyclic Dipeptide ◽  
Synthetic Compound ◽  
Streptomyces Species

Streptomyces is the most frequently described genus of Actinomycetes, a producer of biologically active secondary metabolites. Indeed, the Streptomyces species produces about 70% of antibiotics and 60% of antifungal molecules used in agriculture. Our study was carried out with the goal of isolating and identifying antimicrobial secondary metabolites from Streptomyces misionensis V16R3Y1 isolated from the date palm rhizosphere (southern Tunisia). This strain presented a broad range of antifungal activity against Fusarium oxysporum, Aspergillus flavus, Penicillium expansum, Aspergillus niger, Candida albicans, Candida metapsilosis, and Candida parapsilosis and antibacterial activity against human pathogenic bacteria, including Escherichia fergusonii, Staphylococcus aureus, Salmonella enterica, Enterococcus faecalis, Bacillus cereus and Pseudomonas aeruginosa. The purification procedure entailed ethyl acetate extract, silica gel column, and thin layer chromatography. Based on 1H NMR metabolomic procedure application, also supported by the GC-MS analysis, cyclic dipeptide (l-Leucyl-l-Proline) was identified as the major compound in the bioactive fraction. In order to confirm the identity of the active compound and to have a large quantity thereof, a chemical synthesis of the cyclic dipeptide was performed. The synthetic compound was obtained with a very good yield (50%) and presented almost the same effect compared to the extracted fraction. This study indicates for the first time that Streptomyces misionensis V16R3Y1 exhibits a broad spectrum of antimicrobial activities, produced cyclic dipeptide (l-Leucyl-l-Proline) and might have potential use as a natural agent for pharmaceutical and agri-food applications.

scholarly journals Binding of aldolase to actin-containing filaments. Evidence of interaction with the regulatory proteins of skeletal muscle

1980 ◽  
Vol 186 (1) ◽  
pp. 89-98 ◽  
Author(s):  
T P Walsh ◽  
D J Winzor ◽  
F M Clarke ◽  
C J Masters ◽  
D J Morton
Keyword(s):  
Skeletal Muscle ◽  
Binding Constant ◽  
Moving Boundary ◽  
Regulatory Protein ◽  
Regulatory Proteins ◽  
Affinity Constant ◽  
Quantitative Studies ◽  
Multiple Binding Sites ◽  
Multiple Binding ◽  
Troponin Complex

The interactions of aldolase with regulatory proteins of rabbit skeletal muscle were investigated by moving-boundary electrophoresis. A salt-dependent interaction of troponin, tropomyosin and the tropomyosin-troponin complex with aldolase was detected, the tropomyosin-troponin complex displaying a greater affinity for the enzyme than did either regulatory protein alone. The results indicate that aldolase possesses multiple binding sites (three or more) for these muscle proteins. Quantitative studies of the binding of aldolase to actin-containing filaments showed the interaction to be influenced markedly by the presence of these muscle regulatory proteins on the filaments. In imidazole/HCl buffer, I 0.088, pH 6.8, aldolase binds to F-actin with an affinity constant of 2 × 10(5) M-1 and a stoicheiometry of one tetrameric aldolase molecule per 14 monomeric actin units. Use of F-actin-tropomyosin as adsorbent results in a doubling of the stoicheiometry without significant change in the intrinsic association constant. With F-actin-tropomyosin-troponin a lower binding constant (6 × 10(4) M-1) but even greater stoicheiometry (4:14 actin units) are observed. The presence of Ca2+ (0.1 mM) decreases this stoicheiometry to 3:14 without affecting significantly the magnitude of the intrinsic binding constant.

scholarly journals GCMS ANALYSIS OF ENDOPHYTIC FUNGI CURVULARIA AERIA MTCC-12847 ISOLATED FROM TRIBULUS TERRESTRIS L.

2019 ◽  
pp. 26-32
Author(s):  
Kamana Sahani ◽  
DEEPENDRA THAKUR
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Secondary Metabolites ◽  
Ethyl Acetate ◽  
Endophytic Fungi ◽  
Active Compound ◽  
Ethyl Acetate Extract ◽  
Gas Chromatography Mass Spectrometry ◽  
Tribulus Terrestris ◽  
Chromatography Mass Spectrometry

Objective: The objective of the present investigation was to perform the Gas Chromatography-mass spectrometry (GCMS) analysis of endophytic fungi Curvularia aeria MTCC-12847 isolated from Tribulus terrestris L. to find out the active compound present in the extract. Methods: The endophytic fungi were isolated from the plant Tribulus Terrestris L., leaf which was cultivated in optimized media for the production of secondary metabolites and was extracted using ethyl acetate. Ethyl acetate extract was used for the Gas Chromatography-mass spectrometry (GCMS) analysis. Results: GC-MS analysis of ethyl acetate extract of endophytic fungi revealed the presence of various secondary metabolites, the highest amount present was Palmitic acid (24.54%) and Lowest was Dimethyl 1-phenyl-7-methyl-1-hydroxy-1,4-dihydronaphthalene-2,3-dicarboxylate (5.76%). Conclusion: The endophytic fungal Curvularia aeria MTCC-12847 extract isolated from the Tribulus terrestris L. shows the presence of various bioactive compounds.

scholarly journals Proteomic Insights into Starvation of Nitrogen-Replete Cells of Nostoc sp. PCC 7120 under β-N-Methylamino-L-Alanine (BMAA) Treatment

Toxins ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 372
Author(s):  
Olga A. Koksharova ◽  
Ivan O. Butenko ◽  
Olga V. Pobeguts ◽  
Nina A. Safronova ◽  
Vadim M. Govorun
Keyword(s):  
Molecular Mechanisms ◽  
Nitrogen Starvation ◽  
Regulatory Protein ◽  
Nitrogen Deficiency ◽  
Regulatory Proteins ◽  
Pii Protein ◽  
Significant Difference ◽  
Pcc 7120 ◽  
Nitrogen Excess ◽  
Starvation Conditions

All cyanobacteria produce a neurotoxic non-protein amino acid β-N-methylamino-L-alanine (BMAA). However, the biological function of BMAA in the regulation of cyanobacteria metabolism still remains undetermined. It is known that BMAA suppresses the formation of heterocysts in diazotrophic cyanobacteria under nitrogen starvation conditions, and BMAA induces the formation of heterocyst-like cells under nitrogen excess conditions, by causing the expression of heterocyst-specific genes that are usually “silent” under nitrogen-replete conditions, as if these bacteria receive a nitrogen deficiency intracellular molecular signal. In order to find out the molecular mechanisms underlying this unexpected BMAA effect, we studied the proteome of cyanobacterium Nostoc sp. PCC 7120 grown under BMAA treatment in nitrogen-replete medium. Experiments were performed in two experimental settings: (1) in control samples consisted of cells grown without the BMAA treatment and (2) the treated samples consisted of cells grown with addition of an aqueous solution of BMAA (20 µM). In total, 1567 different proteins of Nostoc sp. PCC 7120 were identified by LC-MS/MS spectrometry. Among them, 80 proteins belonging to different functional categories were chosen for further functional analysis and interpretation of obtained proteomic data. Here, we provide the evidence that a pleiotropic regulatory effect of BMAA on the proteome of cyanobacterium was largely different under conditions of nitrogen-excess compared to its effect under nitrogen starvation conditions (that was studied in our previous work). The most significant difference in proteome expression between the BMAA-treated and untreated samples under different growth conditions was detected in key regulatory protein PII (GlnB). BMAA downregulates protein PII in nitrogen-starved cells and upregulates this protein in nitrogen-replete conditions. PII protein is a key signal transduction protein and the change in its regulation leads to the change of many other regulatory proteins, including different transcriptional factors, enzymes and transporters. Complex changes in key metabolic and regulatory proteins (RbcL, RbcS, Rca, CmpA, GltS, NodM, thioredoxin 1, RpbD, ClpP, MinD, RecA, etc.), detected in this experimental study, could be a reason for the appearance of the “starvation” state in nitrogen-replete conditions in the presence of BMAA. In addition, 15 proteins identified in this study are encoded by genes, which are under the control of NtcA—a global transcriptional regulator—one of the main protein partners and transcriptional regulators of PII protein. Thereby, this proteomic study gives a possible explanation of cyanobacterium starvation under nitrogen-replete conditions and BMAA treatment. It allows to take a closer look at the regulation of cyanobacteria metabolism affected by this cyanotoxin.

scholarly journals Mutations in Transcriptional Regulators Allow Selective Engineering of Signal Integration Logic

mBio ◽  
2014 ◽  
Vol 5 (3) ◽  
Author(s):  
Szabolcs Semsey
Keyword(s):  
Gene Regulatory Networks ◽  
Regulatory Networks ◽  
Regulatory Protein ◽  
Point Mutations ◽  
Building Blocks ◽  
Regulatory Elements ◽  
Regulatory Proteins ◽  
Single Amino Acid ◽  
Signal Integration ◽  
Gene Regulatory

ABSTRACT Bacterial cells monitor their environment by sensing a set of signals. Typically, these environmental signals affect promoter activities by altering the activity of transcription regulatory proteins. Promoters are often regulated by more than one regulatory protein, and in these cases the relevant signals are integrated by certain logic. In this work, we study how single amino acid substitutions in a regulatory protein (GalR) affect transcriptional regulation and signal integration logic at a set of engineered promoters. Our results suggest that point mutations in regulatory genes allow independent evolution of regulatory logic at different promoters. IMPORTANCE Gene regulatory networks are built from simple building blocks, such as promoters, transcription regulatory proteins, and their binding sites on DNA. Many promoters are regulated by more than one regulatory input. In these cases, the inputs are integrated and allow transcription only in certain combinations of input signals. Gene regulatory networks can be easily rewired, because the function of cis-regulatory elements and promoters can be altered by point mutations. In this work, we tested how point mutations in transcription regulatory proteins can affect signal integration logic. We found that such mutations allow context-dependent engineering of signal integration logic at promoters, further contributing to the plasticity of gene regulatory networks.

scholarly journals Platelet activating factor and U44069 stimulate a GTPase activity in human platelets which is distinct from the guanine nucleotide regulatory proteins, Ns and Ni

1986 ◽  
Vol 234 (3) ◽  
pp. 737-740 ◽  
Author(s):  
M D Houslay ◽  
D Bojanic ◽  
A Wilson
Keyword(s):  
Regulatory Protein ◽  
Platelet Activating Factor ◽  
Phospholipid Metabolism ◽  
Human Platelets ◽  
Regulatory Proteins ◽  
Guanine Nucleotide ◽  
Gtpase Activity ◽  
Guanine Nucleotide Regulatory Proteins ◽  
Gtpase Activation ◽  
Guanine Nucleotide Regulatory Protein

Platelet-activating factor (PAF, 2-acetyl-1-alkyl-sn-glycero-3-phosphocholine) and the stable thromboxane-receptor agonist U44069 (9 alpha, 11 beta-epoxymethanoprostaglandin H2) stimulated GTPase activity in platelet membranes in a dose-dependent fashion, yielding Ka values of 12 nM and 27 nM respectively. The degree of GTPase activation elicited by these agents was found to be additive with the GTPase activation due to either the stimulatory (Ns) or inhibitory (Ni) guanine nucleotide regulatory proteins when activated by prostaglandin E1 and adrenaline (+propranolol) respectively. Treatment of membranes with either cholera or pertussis toxins, which inhibited markedly the receptor-mediated stimulation of the GTPase activities of Ns and Ni respectively, had no or only a small effect, respectively, on the GTPase activity stimulated by PAF and U44069. It is suggested that PAF and U44069, which stimulate inositol phospholipid metabolism in platelets, exert actions through a guanine nucleotide regulatory protein which is distinct from Ns and Ni.

Mechanisms of lithium-vasopressin interaction in rabbit cortical collecting tubule

1987 ◽  
Vol 252 (6) ◽  
pp. F1080-F1087 ◽  
Author(s):  
E. Cogan ◽  
M. Svoboda ◽  
M. Abramow
Keyword(s):  
Water Permeability ◽  
Regulatory Protein ◽  
Competitive Inhibitor ◽  
Inhibitory Effect ◽  
Regulatory Proteins ◽  
Collecting Tubule ◽  
Bordetella Pertussis Toxin ◽  
Guanine Nucleotide Regulatory Protein ◽  
Collecting Tubules ◽  
Stimulation Of

In single cortical collecting tubules (CCT) of the rabbit, guanosine 5'-triphosphate (GTP) increased the arginine vasopressin (AVP)-stimulated adenylate cyclase (AC) by 60% (P less than 0.05). In contrast, guanosine 5' O-(2-thio)-diphosphate (GDP-beta S), a competitive inhibitor of GTP action on the stimulatory guanine regulatory protein (Ns), reduced the AVP-stimulated AC activity by 72% (P less than 0.001), indicating the presence of endogenous GTP in the cells under study. That inhibitory effect was reversed by the addition of GTP to the incubation medium. In isolated perfused CCT, cholera toxin (CT) induced a significant increase in water permeability in the absence of AVP. In contrast, Bordetella pertussis toxin (BPT) did not modify the low AVP-independent water permeability. Lithium, an inhibitor of the hydrosmotic action of AVP, also inhibits the hydrosmotic action of CT by 70% (P less than 0.05) but not that of forskolin. The conclusions of the present study are Ns is required for AVP stimulation of AC in the CCT; Ns is functionally active in this system as evidenced by the hydrosmotic effect of CT; the lack of effect of BPT suggests that the low AVP-independent water permeability in the CCT is not the result of a tonic inhibition of the AC operating through the inhibitory guanine nucleotide regulatory protein; and the inhibition by lithium of the hydrosmotic action of AVP in the CCT appears to involve an interaction with the regulatory proteins (probably Ns) or with their binding to the catalytic unit of AC.

Reduced Serum Levels of Cluster of Differentiation 200 in Alcohol-Dependent Patients

2020 ◽  
Vol 55 (4) ◽  
pp. 391-394
Author(s):  
Abhishek Chaturvedi ◽  
Guruprasad Rao ◽  
Samir Kumar Praharaj ◽  
Kanive Parashiva Guruprasad ◽  
Vivek Pais
Keyword(s):  
Alcohol Dependence ◽  
Alcohol Withdrawal ◽  
Addiction Treatment ◽  
Regulatory Protein ◽  
Serum Levels ◽  
Regulatory Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Cluster Of Differentiation ◽  
Alcohol Dependent

Abstract Aim Chronic alcohol consumption can activate and dysregulate the neuroimmune system which leads to neuroinflammation. Neuroimmune regulatory proteins (NIReg) (e.g. Cluster of Differentiation 200 (CD200)) are the regulators of innate immune response and are responsible for silencing the innate immunity and suppression of inflammation. In this study, we explored the changes of serum levels of CD200 in patients with alcohol dependence at baseline, after one-week alcohol withdrawal and after one-month of alcohol abstinence. Methods Seventeen patients with alcohol dependence admitted for de-addiction treatment and 12 healthy controls were enrolled in the study. Blood samples were collected at baseline, after one-week, and after one-month, and CD200 levels were measured using enzyme-linked immunosorbent assay kit and compared with the healthy controls. Results The serum level of the neuroimmune regulatory protein CD200 in alcohol dependent group (at baseline) was significantly lower compared to healthy controls (p=0.003), and increased after one-week, and one-month period. Conclusion The present study indicates that decrease of CD200 serum levels in alcohol dependent patients and its rise during alcohol withdrawal and abstinence may provide a preliminary evidence of the role of neuroimmune regulatory proteins in neuroadaptation during alcohol withdrawal.

scholarly journals Structural and functional insights into transmembrane AMPA receptor regulatory protein complexes

2019 ◽  
Vol 151 (12) ◽  
pp. 1347-1356 ◽  
Author(s):  
Edward C. Twomey ◽  
Maria V. Yelshanskaya ◽  
Alexander I. Sobolevsky
Keyword(s):  
Protein Complexes ◽  
Regulatory Protein ◽  
Regulatory Proteins ◽  
Specific Gene ◽  
Modular Architecture ◽  
Auxiliary Subunits ◽  
Functional Studies ◽  
Excitatory Neurotransmission ◽  
Synaptic Complexes ◽  
The Brain

Fast excitatory neurotransmission is mediated by the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) subtype of ionotropic glutamate receptor (AMPAR). AMPARs initiate depolarization of the postsynaptic neuron by allowing cations to enter through their ion channel pores in response to binding of the neurotransmitter glutamate. AMPAR function is dramatically affected by auxiliary subunits, which are regulatory proteins that form various complexes with AMPARs throughout the brain. The most well-studied auxiliary subunits are the transmembrane AMPAR regulatory proteins (TARPs), which alter the assembly, trafficking, localization, kinetics, and pharmacology of AMPARs. Recent structural and functional studies of TARPs and the TARP-fold germ cell-specific gene 1-like (GSG1L) subunit have provided important glimpses into how auxiliary subunits regulate the function of synaptic complexes. In this review, we put these recent structures in the context of new functional findings in order to gain insight into the determinants of AMPAR regulation by TARPs. We thus reveal why TARPs display a broad range of effects despite their conserved modular architecture.

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