EMD-27334

Single-particle
3.1 Å
EMD-27334 Deposition: 17/06/2022
Map released: 21/09/2022
Last modified: 12/06/2024
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links

EMD-27334

PI 3-kinase alpha with nanobody 3-142

EMD-27334

Single-particle
3.1 Å
EMD-27334 Deposition: 17/06/2022
Map released: 21/09/2022
Last modified: 12/06/2024
Overview 3D View Sample Experiment Validation Volume Browser Additional data Links
Sample Organism: Homo sapiens
Sample: Human PI 3-kinase alpha complex composed of p110alpha and p85alpha with nanobody 3-142 bound
Fitted models: 8dd4 (Avg. Q-score: 0.42)

Deposition Authors: Hart JR , Liu X, Pan C, Liang A, Ueno L , Xu Y, Quezada A, Zou X , Yang S, Zhou Q , Schoonooghe S, Hassanzadeh-Ghassabeh G, Xia T, Shui W, Yang D , Vogt PK , Wang M-W
Nanobodies and chemical cross-links advance the structural and functional analysis of PI3K alpha.
Hart JR , Liu X, Pan C, Liang A, Ueno L , Xu Y, Quezada A, Zou X , Yang S, Zhou Q , Schoonooghe S, Hassanzadeh-Ghassabeh G, Xia T, Shui W, Yang D , Vogt PK , Wang MW
(2022) PNAS , 119 , e2210769119 - e2210769119
PUBMED: 36095215
DOI: doi:10.1073/pnas.2210769119
ISSN: 1091-6490
ASTM: PNASA6
Abstract:
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.