7PPO

Structure of SidJ/CaM bound to SdeA in pre-glutamylation state

7PPO の概要

• エントリーDOI: 10.2210/pdb7ppo/pdb
• EMDBエントリー: 13583
• 分子名称: Ubiquitinating/deubiquitinating enzyme SdeA, Calmodulin-dependent glutamylase SidJ, Calmodulin, ... (5 entities in total)
• 機能のキーワード: glutamylase, pseudokinase, phosphoribosyl, ubiquitination, complex, ligase
• 由来する生物種: Legionella pneumophila; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 222157.76

構造登録者

Adams, M.,Bhogaraju, S. (登録日: 2021-09-14, 公開日: 2021-10-27, 最終更新日: 2021-11-17)

主引用文献

Adams, M.,Sharma, R.,Colby, T.,Weis, F.,Matic, I.,Bhogaraju, S.
Structural basis for protein glutamylation by the Legionella pseudokinase SidJ.
Nat Commun, 12:6174-6174, 2021

PubMed Abstract: Legionella pneumophila (LP) avoids phagocytosis by secreting nearly 300 effector proteins into the host cytosol. SidE family of effectors (SdeA, SdeB, SdeC and SidE) employ phosphoribosyl ubiquitination to target multiple host Rab GTPases and innate immune factors. To suppress the deleterious toxicity of SidE enzymes in a timely manner, LP employs a metaeffector named SidJ. Upon activation by host Calmodulin (CaM), SidJ executes an ATP-dependent glutamylation to modify the catalytic residue Glu860 in the mono-ADP-ribosyl transferase (mART) domain of SdeA. SidJ is a unique glutamylase that adopts a kinase-like fold but contains two nucleotide-binding pockets. There is a lack of consensus about the substrate recognition and catalytic mechanism of SidJ. Here, we determined the cryo-EM structure of SidJ in complex with its substrate SdeA in two different states of catalysis. Our structures reveal that both phosphodiesterase (PDE) and mART domains of SdeA make extensive contacts with SidJ. In the pre-glutamylation state structure of the SidJ-SdeA complex, adenylylated E860 of SdeA is inserted into the non-canonical (migrated) nucleotide-binding pocket of SidJ. Structure-based mutational analysis indicates that SidJ employs its migrated pocket for the glutamylation of SdeA. Finally, using mass spectrometry, we identified several transient autoAMPylation sites close to both the catalytic pockets of SidJ. Our data provide unique insights into the substrate recognition and the mechanism of protein glutamylation by the pseudokinase SidJ.

PubMed: 34702826
DOI: 10.1038/s41467-021-26429-y

実験手法

ELECTRON MICROSCOPY (2.91 Å)