Characterization of therapeutic targets from Aspergillus fumigates in response to Adjunctive Combination therapy (Ketoconazole with EDTA).

2021 ◽  
Vol 19 ◽  
Author(s):  
Sonam Ruhil ◽  
Vikash Kumar ◽  
Monika Malik ◽  
Meenakshi Balhara ◽  
Anil Kumar Chhillar
Keyword(s):  
Combination Therapy ◽  
Therapeutic Targets ◽  
Major Facilitator Superfamily ◽  
Fungal Resistance ◽  
Sublethal Dose ◽  
Fungal Virulence ◽  
Peptide Mass ◽  
Proteomic Approach ◽  
Aspergillus Fumigates ◽  
Haemolytic Assay

Background: The Kingdom (Fungi) comprises numerous species that are associated with numerous fungal diseases. Moreover, the fungal resistance, stagnation in the development of antifungal agents and unacceptably high mortality rate associated with some resistant fungus indicates that alternative therapeutic options should consider. Objective: The objective of this study was to find out new therapeutic targets of A.fumigatus in response to adjunctive combination i.e. Ketoconazole (KTZ) plus EDTA. Methods: A.fumigatus was cultured in absence and presence of sublethal dose (MIC 50) of EDTA, KTZ and Combination of KTZ plus EDTA. The cytosolic proteins were extracted by mechanical grinding of fungal cells.The protein profile was studied by using proteomic approach and identification of protein was done by MALDI-TOF/MS. The morphologicaleffect of combination on A. fumigatus was studied by Scanning Electron Microscopy (SEM) and toxic effect on erythrocytes by haemolytic assay. Result: The combination of KTZ with EDTA was non-toxic upto 500 µg/ml by MTT assay. It inhibits the expression offollowing proteins-Glutamatedehydrogenase, Phenyl alanyl t-RNA synhetase POD G, CaO19-5601, AN6454.2 (Conserved domain; MFS (Major Facilitator Superfamily), serine/threonine protein kinase and dipeptidyl peptidase (identified by peptide mass finger printing).Some of these proteins are involved in hyphal development. Morphological defects on development of fungus (like disrupted hyphal tips,phialide) were observed. Conclusion: These targets can be used for novel drug development as some of them are involved in fungal virulence and adjunctive combination therapy can be an optimistic approach.

scholarly journals Comparative Secretome Analyses of Three Bacillus anthracis Strains with Variant Plasmid Contents

2005 ◽  
Vol 73 (6) ◽  
pp. 3646-3658 ◽  
Author(s):  
Janine M. Lamonica ◽  
MaryAnn Wagner ◽  
Michel Eschenbrenner ◽  
Leanne E. Williams ◽  
Tabbi L. Miller ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Protective Antigen ◽  
Isocitrate Lyase ◽  
Lethal Factor ◽  
Protein A ◽  
Surface Protein ◽  
Computer Assisted ◽  
Two Dimensional Gel Electrophoresis ◽  
Peptide Mass ◽  
Proteomic Approach

ABSTRACT Bacillus anthracis, the causative agent of anthrax, secretes numerous proteins into the extracellular environment during infection. A comparative proteomic approach was employed to elucidate the differences among the extracellular proteomes (secretomes) of three isogenic strains of B. anthracis that differed solely in their plasmid contents. The strains utilized were the wild-type virulent B. anthracis RA3 (pXO1+ pXO2+) and its two nonpathogenic derivative strains: the toxigenic, nonencapsulated RA3R (pXO1+ pXO2−) and the totally cured, nontoxigenic, nonencapsulated RA3:00 (pXO1− pXO2−). Comparative proteomics using two-dimensional gel electrophoresis followed by computer-assisted gel image analysis was performed to reveal unique, up-regulated, or down-regulated secretome proteins among the strains. In total, 57 protein spots, representing 26 different proteins encoded on the chromosome or pXO1, were identified by peptide mass fingerprinting. S-layer-derived proteins, such as Sap and EA1, were most frequently observed. Many sporulation-associated enzymes were found to be overexpressed in strains containing pXO1+. This study also provides evidence that pXO2 is necessary for the maximal expression of the pXO1-encoded toxins lethal factor (LF), edema factor (EF), and protective antigen (PA). Several newly identified putative virulence factors were observed; these include enolase, a high-affinity zinc uptake transporter, the peroxide stress-related alkyl hydroperoxide reductase, isocitrate lyase, and the cell surface protein A.

A comparative proteomic approach to analyse structure, function and evolution of rice chitinases: a step towards increasing plant fungal resistance

2012 ◽  
Vol 18 (11) ◽  
pp. 4761-4780 ◽  
Author(s):  
Kishore Sarma ◽  
Budheswar Dehury ◽  
Jagajjit Sahu ◽  
Ranjan Sarmah ◽  
Smita Sahoo ◽  
...  
Keyword(s):  
Structure Function ◽  
Fungal Resistance ◽  
Proteomic Approach

scholarly journals Revealing the Therapeutic Targets and Mechanism of Baicalin for Anti-chronic Gastritis Through TMT Proteomic Approach

2021 ◽  
Author(s):  
Wanli Ji ◽  
Yan Huo ◽  
Yu Zhang ◽  
Xinhong Wang ◽  
Yifan Zhang
Keyword(s):  
Chronic Gastritis ◽  
Therapeutic Targets ◽  
Interaction Network ◽  
B Cell Lymphoma ◽  
Treated Group ◽  
Sprague Dawley ◽  
Protein Protein Interaction ◽  
Proteomic Approach ◽  
Proteome Changes ◽  
Factor Α

Abstract BackgroundBaicalin (BCL) is a natural compound with beneficial activities, including antioxidant, anti-inflammatory and immunomodulatory. To investigate the therapeutic action of baicalin treatment in ethanol-induced chronic gastritis. Here, we investigated the proteome changes in the gastric tissue to elucidate the therapeutic targets of baicalin in chronic gastritis by TMT-based quantitative proteomics.ResultsUsing TMT-based quantitative proteomics, a total of the 4,452 proteins were identified and quantified in the gastric antrum tissue of Sprague–Dawley rats. Of these, 107 differentially expressed proteins, including 44 up-regulated and 63 down-regulated proteins, were uncovered in the baicalin-treated group as compared with the untreated group with ethanol-induced gastritis. Furthermore, the expression of TPM2, GIMAP4, and Mpc1 was validated using Western Blot. Baicalin could decrease the production of interleukin (IL)-2, IL-8 and tumor necrosis factor-α (TNF-α), while increase the expression of epidermal growth factor (EGF) and B-cell lymphoma-2 (Bcl-2). Notably, protein-protein interaction network analysis revealed the widespread interactions mediated by baicalin.ConclusionsWe investigated the effects and potential mechanism of baicalin in chronic gastritis. Proteomic technology was used to explore baicalin-affected proteins and some signaling pathways. The results may provide important insights into the discovery of potential target proteins for the treatment of chronic gastritis.

scholarly journals 2,3-Dihydroxypropane-1-sulfonate degraded by Cupriavidus pinatubonensis JMP134: purification of dihydroxypropanesulfonate 3-dehydrogenase

Microbiology ◽  
2010 ◽  
Vol 156 (5) ◽  
pp. 1556-1564 ◽  
Author(s):  
Jutta Mayer ◽  
Thomas Huhn ◽  
Michael Habeck ◽  
Karin Denger ◽  
Klaus Hollemeyer ◽  
...  
Keyword(s):  
Ralstonia Eutropha ◽  
Peptide Mass Fingerprinting ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Major Facilitator Superfamily ◽  
Bacterial Degradation ◽  
Uptake System ◽  
Peptide Mass ◽  
Terrestrial Environments ◽  
Sulfoacetaldehyde Acetyltransferase

2,3-Dihydroxypropane-1-sulfonate (DHPS) is a widespread intermediate in plant and algal transformations of sulfoquinovose (SQ) from the plant sulfolipid sulfoquinovosyl diacylglycerol. Further, DHPS is recovered quantitatively during bacterial degradation of SQ by Klebsiella sp. strain ABR11. DHPS is also a putative precursor of sulfolactate in e.g. Ruegeria pomeroyi DSS-3. A bioinformatic approach indicated that some 28 organisms with sequenced genomes might degrade DHPS inducibly via sulfolactate, with three different desulfonative enzymes involved in its degradation in different organisms. The hypothesis for Cupriavidus pinatubonensis JMP134 (formerly Ralstonia eutropha) involved a seven-gene cluster (Reut_C6093–C6087) comprising a LacI-type transcriptional regulator, HpsR, a major facilitator superfamily uptake system, HpsU, three NAD(P)+-coupled DHPS dehydrogenases, HpsNOP, and (R)-sulfolactate sulfo-lyase (SuyAB) [EC 4.4.1.24]. HpsOP effected a DHPS-racemase activity, and HpsN oxidized (R)-DHPS to (R)-sulfolactate. The hypothesis for Roseovarius nubinhibens ISM was similar, but involved a tripartite ATP-independent transport system for DHPS, HpsKLM, and two different desulfonative enzymes, (S)-cysteate sulfo-lyase [EC 4.4.1.25] and sulfoacetaldehyde acetyltransferase (Xsc) [EC 2.3.3.15]. Representative organisms were found to grow with DHPS and release sulfate. C. pinatubonensis JMP134 was found to express at least one NAD(P)+-coupled DHPS dehydrogenase inducibly, and three different peaks of activity were separated by anion-exchange chromatography. Protein bands (SDS-PAGE) were subjected to peptide-mass fingerprinting, which identified the corresponding genes (hpsNOP). Purified HpsN converted DHPS to sulfolactate. Reverse-transcription PCR confirmed that hpsNOUP were transcribed inducibly in strain JMP134, and that hpsKLM and hpsNOP were transcribed in strain ISM. DHPS degradation is widespread and diverse, implying that DHPS is common in marine and terrestrial environments.

scholarly journals Hyperoxia-Induced Protein Alterations in Renal Rat Tissue: A Quantitative Proteomic Approach to Identify Hyperoxia-Induced Effects in Cellular Signaling Pathways

2015 ◽  
Vol 2015 ◽  
pp. 1-12 ◽  
Author(s):  
Jochen Hinkelbein ◽  
Lennert Böhm ◽  
Oliver Spelten ◽  
David Sander ◽  
Stefan Soltész ◽  
...  
Keyword(s):  
Protein Expression ◽  
Rat Kidney ◽  
Peptide Mass Fingerprinting ◽  
Renal Tissue ◽  
Short Term ◽  
Two Dimensional Gel Electrophoresis ◽  
Normobaric Hyperoxia ◽  
Peptide Mass ◽  
Proteomic Approach ◽  
Hyperoxia Exposure

Introduction. In renal tissue as well as in other organs, supranormal oxygen pressure may lead to deleterious consequences on a cellular level. Additionally, hyperoxia-induced effect in cells and related free radicals may potentially contribute to renal failure. The aim of this study was to analyze time-dependent alterations of rat kidney protein expression after short-term normobaric hyperoxia using proteomics and bioinformatic approaches.Material and Methods.N=36Wistar rats were randomized into six different groups: three groups with normobaric hyperoxia (exposure to 100% oxygen for 3 h) and three groups with normobaric normoxia (NN; room air). After hyperoxia exposure, kidneys were removed immediately, after 3 days and after 7 days. Kidney lysates were analyzed by two-dimensional gel electrophoresis followed by peptide mass fingerprinting using tandem mass spectrometry. Statistical analysis was performed with DeCyder 2D software (p<0.01). Biological functions of differential regulated proteins were studied using functional network analysis (Ingenuity Pathways Analysis and PathwayStudio).Results. Expression of 14 proteins was significantly altered(p<0.01): eight proteins (MEP1A_RAT, RSSA_RAT, F16P1_RAT, STML2_RAT, BPNT1_RAT, LGMN_RAT, ATPA_RAT, and VDAC1_RAT) were downregulated and six proteins (MTUS1_RAT, F16P1_RAT, ACTG_RAT, ACTB_RAT, 2ABA_RAT, and RAB1A_RAT) were upregulated. Bioinformatic analyses revealed an association of regulated proteins with inflammation.Conclusions. Significant alterations in renal protein expression could be demonstrated for up to 7 days even after short-term hyperoxia. The identified proteins indicate an association with inflammation signaling cascades. MEP1A and VDAC1 could be promising candidates to identify hyperoxic injury in kidney cells.

scholarly journals Proteomic signatures of 16 major types of human cancer reveal universal and cancer-type-specific proteins for the identification of potential therapeutic targets

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Yangying Zhou ◽  
T. Mamie Lih ◽  
Jianbo Pan ◽  
Naseruddin Höti ◽  
Mingming Dong ◽  
...  
Keyword(s):  
Large Scale ◽  
Human Cancer ◽  
Therapeutic Targets ◽  
Small Cell ◽  
Tissue Type ◽  
Cancer Type ◽  
Proteomic Profiling ◽  
Human Cancers ◽  
Proteomic Approach ◽  
Specific Proteins

Abstract Background Proteomic characterization of cancers is essential for a comprehensive understanding of key molecular aberrations. However, proteomic profiling of a large cohort of cancer tissues is often limited by the conventional approaches. Methods We present a proteomic landscape of 16 major types of human cancer, based on the analysis of 126 treatment-naïve primary tumor tissues, 94 tumor-matched normal adjacent tissues, and 12 normal tissues, using mass spectrometry-based data-independent acquisition approach. Results In our study, a total of 8527 proteins were mapped to brain, head and neck, breast, lung (both small cell and non-small cell lung cancers), esophagus, stomach, pancreas, liver, colon, kidney, bladder, prostate, uterus and ovary cancers, including 2458 tissue-enriched proteins. Our DIA-based proteomic approach has characterized major human cancers and identified universally expressed proteins as well as tissue-type-specific and cancer-type-specific proteins. In addition, 1139 therapeutic targetable proteins and 21 cancer/testis (CT) antigens were observed. Conclusions Our discoveries not only advance our understanding of human cancers, but also have implications for the design of future large-scale cancer proteomic studies to assist the development of diagnostic and/or therapeutic targets in multiple cancers.

scholarly journals PdMFS1 Transporter Contributes to Penicilliun digitatum Fungicide Resistance and Fungal Virulence during Citrus Fruit Infection

2019 ◽  
Vol 5 (4) ◽  
pp. 100 ◽  
Author(s):  
Marta de Ramón-Carbonell ◽  
Mario López-Pérez ◽  
Luis González-Candelas ◽  
Paloma Sánchez-Torres
Keyword(s):  
Fungicide Resistance ◽  
Citrus Fruit ◽  
Major Facilitator Superfamily ◽  
Fungal Virulence ◽  
Disease Symptoms ◽  
Knock Out ◽  
Wide Range ◽  
Major Facilitator ◽  
Chemical Fungicides ◽  
Mfs Transporter

A new Penicillium digitatum major facilitator superfamily (MFS) transporter (PdMFS1) was identified and functionally characterized in order to shed more light on the mechanisms underlying fungicide resistance. PdMFS1 can play an important role in the intensification of resistance to fungicides normally used in P. digitatum postharvest treatments. In the PdMFS1 disrupted mutants, a slight effect in response to chemical fungicides was observed, but fungicide sensitivity was highly affected in the overexpression mutants which became resistant to wide range of chemical fungicides. Moreover, P. digitatum knock-out mutants exhibited a lower rate of fungal virulence when infected oranges were stored at 20 °C. Disease symptoms were higher in the PdMFS1 overexpression mutants coming from the low-virulent P. digitatum parental strain. In addition, the gene expression analysis showed an induction of PdMFS1 transcription in all overexpression mutants regardless from which progenitor came from, and four-time intensification of the parental wild type strain during citrus infection reinforcing PdMFS1 role in fungal virulence. The P. digitatum MFS transporter PdMFS1 contributes not only to the acquisition of wide range of fungicide resistance but also in fungal virulence during citrus infection.

scholarly journals Comparative Proteomic Approach Identifies Pkm2 and Cofilin-1 as Potential Diagnostic, Prognostic and Therapeutic Targets for Pulmonary Adenocarcinoma

PLoS ONE ◽  
2011 ◽  
Vol 6 (11) ◽  
pp. e27309 ◽  
Author(s):  
Xing-chen Peng ◽  
Feng-ming Gong ◽  
Yu-wei Zhao ◽  
Liang-xue Zhou ◽  
Ying-wei Xie ◽  
...  
Keyword(s):  
Therapeutic Targets ◽  
Pulmonary Adenocarcinoma ◽  
Proteomic Approach

scholarly journals Lipid Flippase Subunit Cdc50 Mediates Drug Resistance and Virulence inCryptococcus neoformans

mBio ◽  
2016 ◽  
Vol 7 (3) ◽  
Author(s):  
Wei Huang ◽  
Guojian Liao ◽  
Gregory M. Baker ◽  
Yina Wang ◽  
Richard Lau ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Cryptococcus Neoformans ◽  
Fungal Pathogen ◽  
Treatment Options ◽  
Membrane Lipid ◽  
Fungal Resistance ◽  
Fungal Virulence ◽  
Innate Resistance ◽  
Fungal Meningitis ◽  
Lipid Flippase

ABSTRACTCryptococcus neoformansis a human fungal pathogen and a major cause of fungal meningitis in immunocompromised individuals. Treatment options for cryptococcosis are limited. Of the two major antifungal drug classes, azoles are active againstC. neoformansbut exert a fungistatic effect, necessitating long treatment regimens and leaving open an avenue for emergence of azole resistance. Drugs of the echinocandin class, which target the glucan synthase and are fungicidal against a number of other fungal pathogens, such asCandidaspecies, are ineffective againstC. neoformans. Despite the sensitivity of the target enzyme to the drug, the reasons for the innate resistance ofC. neoformansto echinocandins remain unknown. To understand the mechanism of echinocandin resistance inC. neoformans, we screened gene disruption and gene deletion libraries for mutants sensitive to the echinocandin-class drug caspofungin and identified a mutation ofCDC50, which encodes the β-subunit of membrane lipid flippase. We found that the Cdc50 protein localized to membranes and that its absence led to plasma membrane defects and enhanced caspofungin penetration into the cell, potentially explaining the increased caspofungin sensitivity. Loss ofCDC50also led to hypersensitivity to the azole-class drug fluconazole. Interestingly, in addition to functioning in drug resistance,CDC50was also essential for fungal resistance to macrophage killing and for virulence in a murine model of cryptococcosis. Furthermore, the surface ofcdc50Δcells contained increased levels of phosphatidylserine, which has been proposed to act as a macrophage recognition signal. Together, these results reveal a previously unappreciated role of membrane lipid flippase inC. neoformansdrug resistance and virulence.IMPORTANCECryptococcus neoformansis a fungal pathogen that is the most common cause of fungal meningitis, causing over 620,000 deaths annually. The treatment options for cryptococcosis are very limited. The most commonly used drugs are either fungistatic (azoles) or highly toxic (amphotericin B). Echinocandins are the newest fungicidal drug class that works well in treating candidiasis and aspergillosis, yet they are ineffective in treating cryptococcosis. In this study, we showed that the regulatory subunit of the lipid translocase (flippase), a protein that regulates the asymmetrical orientation of membrane lipids, is required forC. neoformansresistance to caspofungin, as well as for virulence during infection. This discovery identifies lipid flippase as a potentialC. neoformansdrug target, which plays an important role in the innate resistance ofC. neoformansto echinocandins and in fungal virulence.

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