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{"title"=>"Chemical Inhibition of Bacterial Protein Tyrosine Phosphatase Suppresses Capsule Production", "type"=>"journal", "authors"=>[{"first_name"=>"Alistair J.", "last_name"=>"Standish"}, {"first_name"=>"Angela A.", "last_name"=>"Salim"}, {"first_name"=>"Hua", "last_name"=>"Zhang"}, {"first_name"=>"Robert J.", "last_name"=>"Capon"}, {"first_name"=>"Renato", "last_name"=>"Morona"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"22629313", "issn"=>"1932-6203", "doi"=>"10.1371/journal.pone.0036312", "pui"=>"366356961", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-84866917514", "sgr"=>"84866917514"}, "id"=>"534e416a-9db6-3f19-a4f6-b195a297e045", "abstract"=>"Capsule polysaccharide is a major virulence factor for a wide range of bacterial pathogens, including Streptococcus pneumoniae. The biosynthesis of Wzy-dependent capsules in both gram-negative and -positive bacteria is regulated by a system involving a protein tyrosine phosphatase (PTP) and a protein tyrosine kinase. However, how the system functions is still controversial. In Streptococcus pneumoniae, a major human pathogen, the system is present in all but 2 of the 93 serotypes found to date. In order to study this regulation further, we performed a screen to find inhibitors of the phosphatase, CpsB. This led to the observation that a recently discovered marine sponge metabolite, fascioquinol E, inhibited CpsB phosphatase activity both in vitro and in vivo at concentrations that did not affect the growth of the bacteria. This inhibition resulted in decreased capsule synthesis in D39 and Type 1 S. pneumoniae. Furthermore, concentrations of Fascioquinol E that inhibited capsule also lead to increased attachment of pneumococci to a macrophage cell line, suggesting that this compound would inhibit the virulence of the pathogen. Interestingly, this compound also inhibited the phosphatase activity of the structurally unrelated gram-negative PTP, Wzb, which belongs to separate family of protein tyrosine phosphatases. Furthermore, incubation with Klebsiella pneumoniae, which contains a homologous phosphatase, resulted in decreased capsule synthesis. Taken together, these data provide evidence that PTPs are critical for Wzy-dependent capsule production across a spectrum of bacteria, and as such represents a valuable new molecular target for the development of anti-virulence antibacterials.", "link"=>"http://www.mendeley.com/research/chemical-inhibition-bacterial-protein-tyrosine-phosphatase-suppresses-capsule-production", "reader_count"=>34, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>3, "Researcher"=>5, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>2, "Lecturer"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>3, "Researcher"=>5, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>2, "Lecturer"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>5, "Medicine and Dentistry"=>4, "Agricultural and Biological Sciences"=>18, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>2, "Immunology and Microbiology"=>1, "Earth and Planetary Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Chemistry"=>{"Chemistry"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Earth and Planetary Sciences"=>{"Earth and Planetary Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>18}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Environmental Science"=>{"Environmental Science"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"China"=>1}, "group_count"=>1}

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• http://doi.org/10.1128/JB.01545-14
• http://doi.org/10.1016/j.ijmm.2014.06.002
• http://doi.org/10.1016/j.mib.2016.06.001
• http://doi.org/10.1039/C7DT01350G
• http://doi.org/10.1080/1040841X.2016.1249335
• http://doi.org/10.1099/mic.0.000239
• http://doi.org/10.1002/1873-3468.13797
• http://doi.org/10.1128/JB.02106-14
• http://doi.org/10.1016/j.bbrc.2012.11.049
• http://doi.org/10.1099/mic.0.080747-0
• http://doi.org/10.1146/annurev-micro-020415-111342
• http://doi.org/10.1016/j.jmb.2016.06.016
• http://doi.org/10.1128/CMR.00024-15
• http://doi.org/10.3389/fmicb.2020.01596
• http://doi.org/10.1371/journal.pone.0190402
• http://doi.org/10.1111/jpy.12025
• http://doi.org/10.1186/s13049-019-0596-4
• http://doi.org/10.1016/j.lfs.2020.118621
• http://doi.org/10.1089/ars.2013.5726
• http://doi.org/10.1128/JB.00205-19
• http://doi.org/10.1093/femsre/fuaa072
• http://doi.org/10.3390/md11051602
• http://doi.org/10.1039/C5RA14596A

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/638046"], "description"=>"<p>Total CPS preparations were isolated from equal numbers of bacteria after incubation with FQE for 1 h. CPS levels were analysed by either (A) uronic acid assay or alternatively (B) by separating CPS on SDS-PAGE, transferring to Nylon and the probing with α-cps2 as described in the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036312#s4\" target=\"_blank\">materials and methods</a>. Data in (A) is from ≥3 independent experiments (5 µM vs 1.25 µM; * - <i>P</i><0.05 by Student’s <i>t</i>-test).</p>", "links"=>[], "tags"=>["decreases", "capsule", "synthesis"], "article_id"=>308539, "categories"=>["Molecular Biology", "Microbiology"], "users"=>["Alistair J. Standish", "Angela A. Salim", "Hua Zhang", "Robert J. Capon", "Renato Morona"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036312.g004", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FQE_decreases_capsule_synthesis_in_S_pneumoniae_D39_/308539", "title"=>"FQE decreases capsule synthesis in <i>S. pneumoniae</i> D39.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:09:09"}
• {"files"=>["https://ndownloader.figshare.com/files/637863"], "description"=>"<p>(A) Structure of FQE which (B) inhibited CpsB dephosphorlyation of <i>p</i>NPP with IC₅₀ = 5.21 µM. (C) CpsB inhibition of pNPP dephosphorylation by FQE (10 µM) was investigated with increasing concentrations of CpsB^H5H7. Data shown is from three independent experiments (*** - <i>P</i><0.001 by Student’s <i>t</i>-test compared to no addition of CpsB^H5H7).</p>", "links"=>[], "tags"=>["inhibits", "cpsb", "dephosphorylation"], "article_id"=>308361, "categories"=>["Molecular Biology", "Microbiology"], "users"=>["Alistair J. Standish", "Angela A. Salim", "Hua Zhang", "Robert J. Capon", "Renato Morona"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036312.g002", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FQE_inhibits_CpsB_dephosphorylation_of_p_NPP_/308361", "title"=>"FQE inhibits CpsB dephosphorylation of <i>p</i>NPP.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:08:14"}
• {"files"=>["https://ndownloader.figshare.com/files/637948"], "description"=>"<p><i>S. pneumoniae</i> D39 were grown to mid log phase in THY (OD₆₀₀ ≈ 0.35) and FQE at indicated concentrations were added. (A) These concentrations (µM) had no statistically significant effect on CFU/ml after 30, 60 and 120 mins. (B) Whole cell lysates were prepared from these cells, which were separated by SDS-PAGE and analyzed by immunoblotting using anti-CpsD, or anti-phosphotyrosine (to detect CpsD-P). (C) Densitometric analysis of CpsD-P from three separate experiments. The effect with addition of 5 µM was significantly higher than compared with 1.25 µM FQE (* - <i>P</i><0.05 by Student’s <i>t</i>-test). (D) For comparison, the effect of an in-frame <i>cpsB</i> deletion mutant on CpsD-P is shown.</p>", "links"=>[], "tags"=>["increases", "cpsd-p"], "article_id"=>308441, "categories"=>["Molecular Biology", "Microbiology"], "users"=>["Alistair J. Standish", "Angela A. Salim", "Hua Zhang", "Robert J. Capon", "Renato Morona"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036312.g003", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FQE_increases_CpsD_P_in_S_pneumoniae_D39_/308441", "title"=>"FQE increases CpsD-P in <i>S. pneumoniae</i> D39.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:08:37"}
• {"files"=>["https://ndownloader.figshare.com/files/638128"], "description"=>"<p>D39 was incubated with FQE and then assessed for its ability to associate with RAW 264.7 cells as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036312#s4\" target=\"_blank\">Materials and Methods</a>. Data is presented as % association relative to inoculum. D39<i>cpsB</i>? was used for comparison purposes. Results are from three independent experiments (** - P<0.01; * - P<0.05 compared to D39 by Student’s <i>t</i>-test).</p>", "links"=>[], "tags"=>["increases", "attachment", "d39"], "article_id"=>308618, "categories"=>["Molecular Biology", "Microbiology"], "users"=>["Alistair J. Standish", "Angela A. Salim", "Hua Zhang", "Robert J. Capon", "Renato Morona"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036312.g005", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FQE_increases_attachment_of_D39_to_macrophages_/308618", "title"=>"FQE increases attachment of D39 to macrophages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:09:31"}
• {"files"=>["https://ndownloader.figshare.com/files/638183"], "description"=>"<p>(A) His₆Wzb dephosphorylation of <i>p</i>NPP in presence of FQE in 1 M Tris pH 7.0 at 37°C. (B) Total CPS preparations from <i>K. pneumoniae</i> incubated with FQE were analysed by uronic acid assay. Data is from four independent experiments (20 µM vs 5 µM; * - <i>P</i><0.05 by Student’s <i>t</i>-test:).</p>", "links"=>[], "tags"=>["inhibits", "wzb", "cps", "synthesis", "pneumoniae"], "article_id"=>308673, "categories"=>["Molecular Biology", "Microbiology"], "users"=>["Alistair J. Standish", "Angela A. Salim", "Hua Zhang", "Robert J. Capon", "Renato Morona"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036312.g006", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FQE_also_inhibits_E_coli_Wzb_and_CPS_synthesis_in_K_pneumoniae_O1_/308673", "title"=>"FQE also inhibits <i>E. coli</i> Wzb and CPS synthesis in <i>K. pneumoniae O1</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:09:48"}
• {"files"=>["https://ndownloader.figshare.com/files/637798"], "description"=>"<p>The ability of extracts to inhibit His₆CpsB dephosphorylation of <i>p</i>NPP in 1 M Tris pH 8.0 with 1 mM MnCl₂ was investigated in 96 well trays at 37°C. Shown is a plot of the two screening replicates reported as % phosphatase activity relative to the average of particular screening plate. The star represents the extract which produced the pure compound of interest.</p>", "links"=>[], "tags"=>["extract", "inhibitors", "cpsb"], "article_id"=>308287, "categories"=>["Molecular Biology", "Microbiology"], "users"=>["Alistair J. Standish", "Angela A. Salim", "Hua Zhang", "Robert J. Capon", "Renato Morona"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036312.g001", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Screening_of_Marine_Extract_Library_for_inhibitors_of_CpsB_activity_/308287", "title"=>"Screening of Marine Extract Library for inhibitors of CpsB activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:07:51"}

PMC Usage Stats | Further Information

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Relative Metric

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F1000Prime