8EFX
Structure of OtDUB DUB Domain disulfide crosslinked with Ubiquitin
Summary for 8EFX
| Entry DOI | 10.2210/pdb8efx/pdb |
| Descriptor | OtDUB, Ubiquitin (3 entities in total) |
| Functional Keywords | complex, deubiquitinase, hydrolase, hydrolase-signaling protein complex, hydrolase/signaling protein |
| Biological source | Orientia tsutsugamushi More |
| Total number of polymer chains | 2 |
| Total formula weight | 37772.06 |
| Authors | Negron Teron, K.N.,Das, C. (deposition date: 2022-09-09, release date: 2023-09-20, Last modification date: 2024-05-01) |
| Primary citation | Negron Teron, K.I.,Das, C. Cocrystallization of ubiquitin-deubiquitinase complexes through disulfide linkage. Acta Crystallogr D Struct Biol, 79:1044-1055, 2023 Cited by PubMed Abstract: Structural characterization of the recognition of ubiquitin (Ub) by deubiquitinases (DUBs) has largely relied on covalent complexation of the DUB through its catalytic cysteine with a Ub C-terminal electrophile. The Ub electrophiles are accessed through intein chemistry in conjunction with chemical synthesis. Here, it was asked whether DUB-Ub covalent complexes could instead be accessed by simpler disulfide chemistry using a Ub cysteine mutant in which the last glycine has been replaced with a cysteine. The Ub cysteine mutant displayed a wide variability in disulfide formation across a panel of eukaryotic and prokaryotic DUBs, with some showing no detectable reaction while others robustly produced a disulfide complex. Using this approach, two disulfide-linked ubiquitin-bound complexes were crystallized, one involving the Legionella pneumophila effector SdeA DUB and the other involving the Orientia effector OtDUB. These DUBs had previously been crystallized in Ub-bound forms using the C-terminal electrophile strategy and noncovalent complexation, respectively. While the disulfide-linked SdeA DUB-Ub complex crystallized as expected, in the OtDUB complex the disulfide bond to the Ub mutant involved a cysteine that differed from the catalytic cysteine. Disulfide formation with the SdeA DUB catalytic cysteine was accompanied by local distortion of the helix carrying the active-site cysteine, whereas OtDUB reacted with the Ub mutant using a surface-exposed cysteine. PubMed: 37877948DOI: 10.1107/S2059798323008501 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.85 Å) |
Structure validation
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