PXD041657

PXD041657 is an original dataset announced via ProteomeXchange.

Title	Conformational heterogeneity of the BTK PHTH domain drives multiple regulatory states
Description	Full-length BTK has been refractory to structural analysis. The nearest full-length structure of BTK to date consists of the autoinhibited SH3-SH2-kinase core. Precisely how the BTK N-terminal domains (the Pleckstrin homology/Tec homology (PHTH) domain and the long linker that includes proline-rich regions (PRR)) contribute to BTK regulation remains unclear. Here we produce crystals of full-length BTK for the first time. Despite efforts to stabilize the autoinhibited state, the diffraction data still reveals only the SH3-SH2-kinase core with no electron density visible for the PHTH-PRR segment. CryoEM data, on the other hand, provides a glimpse of the PHTH domain. CryoEM reconstructions support conformational heterogeneity in the PHTH-PRR region; the globular PHTH domain adopts a range of states arrayed around the autoinhibited SH3-SH2-kinase core. Upon disassembly of the SH3-SH2-kinase core, an autoinhibitory site on the kinase domain becomes available for PHTH domain binding. This PHTH/kinase autoinhibitory contact is then lost upon interaction of PHTH with PIP3. Membrane-induced dimerization activates BTK and here we solve a structure of an activation loop swapped BTK kinase domain dimer that likely represents the conformational state leading to trans-autophosphorylation. Together, these data provide the first structural insight into full-length BTK and allow a deeper understanding of allosteric control over the BTK kinase domain during distinct stages of activation.
HostingRepository	PRIDE
AnnounceDate	2024-05-24
AnnouncementXML	Submission_2024-05-24_02:10:08.355.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Thomas Wales
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2023-04-19 12:16:13	ID requested
⏵ 1	2024-05-24 02:10:09	announced

10.7554/elife.89489;

Lin DY, Kueffer LE, Juneja P, Wales TE, Engen JR, Andreotti AH, Conformational heterogeneity of the BTK PHTH domain drives multiple regulatory states. Elife, 12():(2024) [pubmed]

submitter keyword: BTK

HDX-MS

hydrogen/deuterium exchange mass spectrometry

Thomas E. Wales
contact affiliation	Department of Chemistry & Chemical Biology, Northeastern University
contact email	t.wales@northeastern.edu
lab head
Thomas Wales
contact affiliation	Northeastern University
contact email	t.wales@northeastern.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD041657
     1. Label: PRIDE project
     2. Name: Conformational heterogeneity of the BTK PHTH domain drives multiple regulatory states