PXD040480 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY |
Description | SPINDLY (SPY) in Arabidopsis thaliana is a novel nucleocytoplasmic protein O-fucosyltransferase (POFUT), which regulates diverse developmental processes. Sequence analysis indicates that AtSPY is distinct from ER-localized POFUTs and contains N-terminal tetratricopeptide-repeats (TPRs) and a C-terminal catalytic domain resembling the O-linked-N-acetylglucosamine (GlcNAc) transferases (OGTs). However, the structural feature that determines the distinct enzymatic selectivity of SPY remains unknown. Here we report the cryo-electron microscopy (cryo-EM) structure of AtSPY and its complex with GDP-fucose, revealing distinct active-site features enabling GDP-fucose instead of UDP-GlcNAc binding. AtSPY forms an antiparallel dimer instead of the X-shaped dimer in human OGT, and its catalytic domain dynamically interconverts among inward, middle, and outward states. The entire eleven TPRs are visible in the middle state, with TPR1 of one subunit touching down on the catalytic domain of the other subunit of the AtSPY dimer, whereas the densities of TPRs 1-5 are absent or severely attenuated in the inward and outward states. Analysis of mass spectrometry, co-IP, in planta fucosylation activity, and cryo-EM data further demonstrates that the N-terminal disordered peptide in SPY contains trans auto-fucosylation sites and inhibits its catalytic activity, whereas TPRs 1-5 regulate SPY activity by interfering with protein substrate binding. |
HostingRepository | PRIDE |
AnnounceDate | 2023-02-28 |
AnnouncementXML | Submission_2023-02-28_07:09:05.164.xml |
DigitalObjectIdentifier | https://dx.doi.org/10.6019/PXD040480 |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Supported dataset by repository |
PrimarySubmitter | JeffreyShabanowitz |
SpeciesList | scientific name: Nicotiana tabacum (Common tobacco); NCBI TaxID: 4097; |
ModificationList | fucosylated residue; monohydroxylated residue; hexosylated residue; iodoacetamide derivatized residue |
Instrument | Orbitrap Fusion |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2023-02-27 13:05:10 | ID requested | |
⏵ 1 | 2023-02-28 07:09:05 | announced | |
Publication List
Keyword List
submitter keyword: SPY, LC-MS, SPINDLY,Protein O-fucosylation |
Contact List
JeffreyShabanowitz |
contact affiliation | University of Virginia |
contact email | js4c@virginia.edu |
lab head | |
JeffreyShabanowitz |
contact affiliation | University of Virginia |
contact email | js4c@virginia.edu |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD040480
- Label: PRIDE project
- Name: Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY