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Visualizing a Dynamic Allosteric Network in Human Herpesvirus Proteases With an Inhibitory Antibody
Visualizing a Dynamic Allosteric Network in Human Herpesvirus Proteases With an Inhibitory Antibody
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20250211153035
- ISBN
- 9798346877233
- DDC
- 574
- 서명/저자
- Visualizing a Dynamic Allosteric Network in Human Herpesvirus Proteases With an Inhibitory Antibody
- 발행사항
- [Sl] : University of California, San Francisco, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 71 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 86-06, Section: B.
- 주기사항
- Advisor: Cheng, Yifan.
- 학위논문주기
- Thesis (Ph.D.)--University of California, San Francisco, 2024.
- 초록/해제
- 요약Exploiting conformational dynamics is a powerful method for inhibiting enzymes and can produce new allosteric inhibitors for managing disease. In this work, we outline the discovery and characterization of an antibody fragment called Fab5 which inhibits the human Cytomegalovirus protease (HCMV Pr). Using ensemble biophysical techniques, we describe the dynamic relationship between Fab5 and HCMV Pr, from which we propose a mechanism of inhibition along with potential insights into the broader conformational network that controls all human Herpesvirus Protease (HHV Pr) activation. Mutagenesis and biochemical characterization allow us to validate our hypotheses in vitro. We provide novel tools and strategies for HHV Pr inhibition that we believe can be leveraged in future therapeutic development. In Chapter 1.1, we use cryogenic electron microscopy (cryo-EM) to reveal the Fab5 binding site on HCMV Pr which we call the Latch Loop. We propose that Fab5 inhibits HCMV Pr by preventing dimerization through an allosteric mechanism that is both mechanistic and dynamic, ultimately preventing the protease from activating. Mutagenic studies demonstrate the previously unrecognized importance of the Latch Loop in the conformational network that links dimerization to HHV Pr activation. In Chapter 1.2, we use X-ray radiolytic Footprinting to survey solvent accessibility changes on HCMV Pr after Fab5 binding. These studies reveal a residue network near the HCMV Pr active site that influences dimerization and, thus, activity despite being buried in the center of the protease.
- 일반주제명
- Biochemistry
- 일반주제명
- Biophysics
- 일반주제명
- Microbiology
- 일반주제명
- Chemistry
- 키워드
- Antibody
- 키워드
- Cytomegalovirus
- 키워드
- Herpes
- 키워드
- Phage-display
- 키워드
- Protease
- 기타저자
- University of California, San Francisco Chemistry and Chemical Biology
- 기본자료저록
- Dissertations Abstracts International. 86-06B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■00520250211153035
■006m o d
■007cr#unu||||||||
■020 ▼a9798346877233
■035 ▼a(MiAaPQ)AAI31637762
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a574
■1001 ▼aZimanyi, Marcell Antal.▼0(orcid)0000-0003-1981-1244
■24510▼aVisualizing a Dynamic Allosteric Network in Human Herpesvirus Proteases With an Inhibitory Antibody
■260 ▼a[Sl]▼bUniversity of California, San Francisco▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a71 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 86-06, Section: B.
■500 ▼aAdvisor: Cheng, Yifan.
■5021 ▼aThesis (Ph.D.)--University of California, San Francisco, 2024.
■520 ▼aExploiting conformational dynamics is a powerful method for inhibiting enzymes and can produce new allosteric inhibitors for managing disease. In this work, we outline the discovery and characterization of an antibody fragment called Fab5 which inhibits the human Cytomegalovirus protease (HCMV Pr). Using ensemble biophysical techniques, we describe the dynamic relationship between Fab5 and HCMV Pr, from which we propose a mechanism of inhibition along with potential insights into the broader conformational network that controls all human Herpesvirus Protease (HHV Pr) activation. Mutagenesis and biochemical characterization allow us to validate our hypotheses in vitro. We provide novel tools and strategies for HHV Pr inhibition that we believe can be leveraged in future therapeutic development. In Chapter 1.1, we use cryogenic electron microscopy (cryo-EM) to reveal the Fab5 binding site on HCMV Pr which we call the Latch Loop. We propose that Fab5 inhibits HCMV Pr by preventing dimerization through an allosteric mechanism that is both mechanistic and dynamic, ultimately preventing the protease from activating. Mutagenic studies demonstrate the previously unrecognized importance of the Latch Loop in the conformational network that links dimerization to HHV Pr activation. In Chapter 1.2, we use X-ray radiolytic Footprinting to survey solvent accessibility changes on HCMV Pr after Fab5 binding. These studies reveal a residue network near the HCMV Pr active site that influences dimerization and, thus, activity despite being buried in the center of the protease.
■590 ▼aSchool code: 0034.
■650 4▼aBiochemistry
■650 4▼aBiophysics
■650 4▼aMicrobiology
■650 4▼aChemistry
■653 ▼aAntibody
■653 ▼aCryogenic electron microscopy
■653 ▼aCytomegalovirus
■653 ▼aHerpes
■653 ▼aPhage-display
■653 ▼aProtease
■690 ▼a0487
■690 ▼a0786
■690 ▼a0410
■690 ▼a0485
■71020▼aUniversity of California, San Francisco▼bChemistry and Chemical Biology.
■7730 ▼tDissertations Abstracts International▼g86-06B.
■790 ▼a0034
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17164721▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
