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High-Throughput Profiling of Sequence Recognition by Phosphotyrosine Signaling Proteins- [electronic resource]
High-Throughput Profiling of Sequence Recognition by Phosphotyrosine Signaling Proteins- [electronic resource]
상세정보
- 자료유형
- 학위논문파일 국외
- 최종처리일시
- 20240214101241
- ISBN
- 9798379793128
- DDC
- 540
- 저자명
- Li, Allyson.
- 서명/저자
- High-Throughput Profiling of Sequence Recognition by Phosphotyrosine Signaling Proteins - [electronic resource]
- 발행사항
- [S.l.]: : Columbia University., 2023
- 발행사항
- Ann Arbor : : ProQuest Dissertations & Theses,, 2023
- 형태사항
- 1 online resource(163 p.)
- 주기사항
- Source: Dissertations Abstracts International, Volume: 85-01, Section: B.
- 주기사항
- Advisor: Shah, Neel H. .
- 학위논문주기
- Thesis (Ph.D.)--Columbia University, 2023.
- 사용제한주기
- This item must not be sold to any third party vendors.
- 초록/해제
- 요약Protein tyrosine kinase and phosphatase domains have binding specificities that depend on the amino acid sequence surrounding the target (phospho)tyrosine residue on their substrates. Although the preferred recognition motifs of many kinase and phosphatase domains have been characterized, we lack a quantitative description of sequence specificity that could guide predictions about signaling pathways or be used to design sequences for biomedical applications. Here, we present a platform that combines genetically-encoded peptide libraries and deep sequencing to profile sequence recognition by tyrosine kinases. We screened several tyrosine kinases against a million-peptide random library and used the resulting profiles to design high-activity sequences and predict phosphorylation efficiencies of substrates. We then screened several kinases against a library containing thousands of human proteome-derived peptides and their naturally-occurring variants. These screens recapitulated independently measured phosphorylation rates and revealed hundreds of phosphosite-proximal mutations that impact phosphosite recognition by tyrosine kinases. Finally, we have made progress towards extending this platform to the analysis of tyrosine phosphatase domains, by optimizing methods to produce tyrosine-phosphorylated bacterial display libraries and implementing methods to detect peptide dephosphorylation on the cell surface. Collectively, these experiments demonstrate the utility of our platform for rapid profiling of sequence specificity by tyrosine kinases and will shed new light on phosphotyrosine signaling.
- 일반주제명
- Chemistry.
- 일반주제명
- Biomedical engineering.
- 일반주제명
- Biochemistry.
- 키워드
- Phosphotyrosine
- 키워드
- Amino acid
- 키워드
- Cell surface
- 기타저자
- Columbia University Chemistry
- 기본자료저록
- Dissertations Abstracts International. 85-01B.
- 기본자료저록
- Dissertation Abstract International
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■00520240214101241
■006m o d
■007cr#unu||||||||
■020 ▼a9798379793128
■035 ▼a(MiAaPQ)AAI30528380
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a540
■1001 ▼aLi, Allyson.
■24510▼aHigh-Throughput Profiling of Sequence Recognition by Phosphotyrosine Signaling Proteins▼h[electronic resource]
■260 ▼a[S.l.]:▼bColumbia University. ▼c2023
■260 1▼aAnn Arbor :▼bProQuest Dissertations & Theses, ▼c2023
■300 ▼a1 online resource(163 p.)
■500 ▼aSource: Dissertations Abstracts International, Volume: 85-01, Section: B.
■500 ▼aAdvisor: Shah, Neel H. .
■5021 ▼aThesis (Ph.D.)--Columbia University, 2023.
■506 ▼aThis item must not be sold to any third party vendors.
■520 ▼aProtein tyrosine kinase and phosphatase domains have binding specificities that depend on the amino acid sequence surrounding the target (phospho)tyrosine residue on their substrates. Although the preferred recognition motifs of many kinase and phosphatase domains have been characterized, we lack a quantitative description of sequence specificity that could guide predictions about signaling pathways or be used to design sequences for biomedical applications. Here, we present a platform that combines genetically-encoded peptide libraries and deep sequencing to profile sequence recognition by tyrosine kinases. We screened several tyrosine kinases against a million-peptide random library and used the resulting profiles to design high-activity sequences and predict phosphorylation efficiencies of substrates. We then screened several kinases against a library containing thousands of human proteome-derived peptides and their naturally-occurring variants. These screens recapitulated independently measured phosphorylation rates and revealed hundreds of phosphosite-proximal mutations that impact phosphosite recognition by tyrosine kinases. Finally, we have made progress towards extending this platform to the analysis of tyrosine phosphatase domains, by optimizing methods to produce tyrosine-phosphorylated bacterial display libraries and implementing methods to detect peptide dephosphorylation on the cell surface. Collectively, these experiments demonstrate the utility of our platform for rapid profiling of sequence specificity by tyrosine kinases and will shed new light on phosphotyrosine signaling.
■590 ▼aSchool code: 0054.
■650 4▼aChemistry.
■650 4▼aBiomedical engineering.
■650 4▼aBiochemistry.
■653 ▼aPhosphotyrosine
■653 ▼aAmino acid
■653 ▼aProtein tyrosine kinase
■653 ▼aPhosphorylation efficiencies
■653 ▼aCell surface
■690 ▼a0485
■690 ▼a0541
■690 ▼a0487
■71020▼aColumbia University▼bChemistry.
■7730 ▼tDissertations Abstracts International▼g85-01B.
■773 ▼tDissertation Abstract International
■790 ▼a0054
■791 ▼aPh.D.
■792 ▼a2023
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T16933389▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
■980 ▼a202402▼f2024
