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TitleNanobodies and chemical cross-links advance the structural and functional analysis of PI3Kα.
Journal, issue, pagesProc Natl Acad Sci U S A, Vol. 119, Issue 38, Page e2210769119, Year 2022
Publish dateSep 20, 2022
AuthorsJonathan R Hart / Xiao Liu / Chen Pan / Anyi Liang / Lynn Ueno / Yingna Xu / Alexandra Quezada / Xinyu Zou / Su Yang / Qingtong Zhou / Steve Schoonooghe / Gholamreza Hassanzadeh-Ghassabeh / Tian Xia / Wenqing Shui / Dehua Yang / Peter K Vogt / Ming-Wei Wang /
PubMed AbstractNanobodies and chemical cross-linking were used to gain information on the identity and positions of flexible domains of PI3Kα. The application of chemical cross-linking mass spectrometry (CXMS) ...Nanobodies and chemical cross-linking were used to gain information on the identity and positions of flexible domains of PI3Kα. The application of chemical cross-linking mass spectrometry (CXMS) facilitated the identification of the p85 domains BH, cSH2, and SH3 as well as their docking positions on the PI3Kα catalytic core. Binding of individual nanobodies to PI3Kα induced activation or inhibition of enzyme activity and caused conformational changes that could be correlated with enzyme function. Binding of nanobody Nb3-126 to the BH domain of p85α substantially improved resolution for parts of the PI3Kα complex, and binding of nanobody Nb3-159 induced a conformation of PI3Kα that is distinct from known PI3Kα structures. The analysis of CXMS data also provided mechanistic insights into the molecular underpinning of the flexibility of PI3Kα.
External linksProc Natl Acad Sci U S A / PubMed:36095215 / PubMed Central
MethodsEM (single particle)
Resolution2.41 - 3.4 Å
Structure data

27327

8dcp

EMDB-27327, PDB-8dcp:
PI 3-kinase alpha with nanobody 3-126
Method: EM (single particle) / Resolution: 2.41 Å

27330

8dcx

EMDB-27330, PDB-8dcx:
PI 3-kinase alpha with nanobody 3-159
Method: EM (single particle) / Resolution: 2.8 Å

27334

8dd4

EMDB-27334, PDB-8dd4:
PI 3-kinase alpha with nanobody 3-142
Method: EM (single particle) / Resolution: 3.1 Å

27336

8dd8

EMDB-27336, PDB-8dd8:
PI 3-kinase alpha with nanobody 3-142, crosslinked with DSG
Method: EM (single particle) / Resolution: 3.4 Å

Chemicals

ChemComp-HOH:
WATER / Water

Source
  • homo sapiens (human)
KeywordsTRANSFERASE / Phosphoinositide 3-kinase (PI3K) / activation / inhibition / nanobody / conformational changes / STRUCTURAL PROTEIN

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