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Development of Bacterial Histidine Kinase Inhibitors and Probes for the Identification of Direct Cellular Targets
Development of Bacterial Histidine Kinase Inhibitors and Probes for the Identification of Direct Cellular Targets
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20260209102833
- ISBN
- 9798293870318
- DDC
- 615
- 서명/저자
- Development of Bacterial Histidine Kinase Inhibitors and Probes for the Identification of Direct Cellular Targets
- 발행사항
- [Sl] : University of Minnesota, 2023
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2023
- 형태사항
- 450 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 87-03, Section: B.
- 주기사항
- Advisor: Carlson, Erin E.
- 학위논문주기
- Thesis (Ph.D.)--University of Minnesota, 2023.
- 초록/해제
- 요약Bacterial resistance to antibiotics is a rapidly increasing threat to human health. New strategies to combat resistant organisms are desperately needed. One potential avenue is targeting two-component systems, the most common bacterial signal transduction pathways that regulate development, metabolism, virulence, and antibiotic resistance. These systems consist of a homodimeric membrane-bound sensor histidine kinase, and a cognate effector, the response regulator. The high sequence conservation in the catalytic and adenosine triphosphate-binding (CA) domain of histidine kinases and their essential role in bacterial signal transduction could enable broad-spectrum antibacterial activity. Through this signal transduction, histidine kinases regulate multiple virulence mechanisms, including toxin production, immune evasion, and antibiotic resistance. Targeting virulence instead of developing bactericidal compounds could reduce evolutionary pressure for acquired resistance.Additionally, compounds targeting the CA domain can potentially impair multiple two-component systems that regulate virulence in one or more pathogens. We conducted structure-activity relationship studies of 2-aminobenzothiazole-based inhibitors designed to target the CA domain of histidine kinases. We found these compounds have anti-virulence activities in Pseudomonas aeruginosa, reducing motility phenotypes and toxin production associated with the pathogenic functions of this bacterium.We devised covalent inhibitors derived from our non-covalent leads using aryl sulfonyl fluoride (aryl SF) as the warhead and a clickable handle to enable subsequent detection of probe-labeled proteins. Aryl SFs modify nucleophilic amino acid side chains but generally react with proteins only when bound in an active site. The resulting bond is slow to hydrolyze under physiologic conditions, making them ideal biological probes. We have determined that these covalent inhibitors have low micromolar activity against HK853. These covalent analogs have also shown potent inhibitions of virulent motility phenotypes in Pseudomonas aeruginosa. The related, clickable probe labels HK853 protein only a single time in whole protein mass spectrometry experiments. Peptide mapping experiments suggest that this probe is labeling a tyrosine residue in the ATP lid of the CA domain, further indicating specific binding. The probe has also shown utility for labeling proteins in vitro and whole-cell experiments for fluorescent gel analysis and in vitro enrichment studies.
- 일반주제명
- Pharmaceutical sciences
- 일반주제명
- Organic chemistry
- 일반주제명
- Microbiology
- 키워드
- Histidine kinase
- 키워드
- Inhibitors
- 키워드
- Virulence
- 기타저자
- University of Minnesota Medicinal Chemistry
- 기본자료저록
- Dissertations Abstracts International. 87-03B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■007cr#unu||||||||
■020 ▼a9798293870318
■035 ▼a(MiAaPQ)AAI30639023
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a615
■1001 ▼aFihn, Conrad Alexander.
■24510▼aDevelopment of Bacterial Histidine Kinase Inhibitors and Probes for the Identification of Direct Cellular Targets
■260 ▼a[Sl]▼bUniversity of Minnesota▼c2023
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2023
■300 ▼a450 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 87-03, Section: B.
■500 ▼aAdvisor: Carlson, Erin E.
■5021 ▼aThesis (Ph.D.)--University of Minnesota, 2023.
■520 ▼aBacterial resistance to antibiotics is a rapidly increasing threat to human health. New strategies to combat resistant organisms are desperately needed. One potential avenue is targeting two-component systems, the most common bacterial signal transduction pathways that regulate development, metabolism, virulence, and antibiotic resistance. These systems consist of a homodimeric membrane-bound sensor histidine kinase, and a cognate effector, the response regulator. The high sequence conservation in the catalytic and adenosine triphosphate-binding (CA) domain of histidine kinases and their essential role in bacterial signal transduction could enable broad-spectrum antibacterial activity. Through this signal transduction, histidine kinases regulate multiple virulence mechanisms, including toxin production, immune evasion, and antibiotic resistance. Targeting virulence instead of developing bactericidal compounds could reduce evolutionary pressure for acquired resistance.Additionally, compounds targeting the CA domain can potentially impair multiple two-component systems that regulate virulence in one or more pathogens. We conducted structure-activity relationship studies of 2-aminobenzothiazole-based inhibitors designed to target the CA domain of histidine kinases. We found these compounds have anti-virulence activities in Pseudomonas aeruginosa, reducing motility phenotypes and toxin production associated with the pathogenic functions of this bacterium.We devised covalent inhibitors derived from our non-covalent leads using aryl sulfonyl fluoride (aryl SF) as the warhead and a clickable handle to enable subsequent detection of probe-labeled proteins. Aryl SFs modify nucleophilic amino acid side chains but generally react with proteins only when bound in an active site. The resulting bond is slow to hydrolyze under physiologic conditions, making them ideal biological probes. We have determined that these covalent inhibitors have low micromolar activity against HK853. These covalent analogs have also shown potent inhibitions of virulent motility phenotypes in Pseudomonas aeruginosa. The related, clickable probe labels HK853 protein only a single time in whole protein mass spectrometry experiments. Peptide mapping experiments suggest that this probe is labeling a tyrosine residue in the ATP lid of the CA domain, further indicating specific binding. The probe has also shown utility for labeling proteins in vitro and whole-cell experiments for fluorescent gel analysis and in vitro enrichment studies.
■590 ▼aSchool code: 0130.
■650 4▼aPharmaceutical sciences
■650 4▼aOrganic chemistry
■650 4▼aMicrobiology
■653 ▼aHistidine kinase
■653 ▼aInhibitors
■653 ▼aPseudomonas aeruginosa
■653 ▼aStructure-activity relationship
■653 ▼aVirulence
■690 ▼a0572
■690 ▼a0490
■690 ▼a0410
■71020▼aUniversity of Minnesota▼bMedicinal Chemistry.
■7730 ▼tDissertations Abstracts International▼g87-03B.
■790 ▼a0130
■791 ▼aPh.D.
■792 ▼a2023
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17365832▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
