6SIU

Crystal structure of IbpAFic2 covalently tethered to Cdc42

6SIU の概要

• エントリーDOI: 10.2210/pdb6siu/pdb
• 分子名称: Protein adenylyltransferase and cysteine protease IbpA, Cell division control protein 42 homolog, SULFATE ION, ... (7 entities in total)
• 機能のキーワード: fic domain, adenosine monophosphate-protein transferase, proximity enabled coupling, transferase
• 由来する生物種: Histophilus somni (strain 2336); 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 116402.80

構造登録者

Gulen, B.,Roselin, M.,Albers, M.,Hedberg, C.,Itzen, A.,Pogenberg, V. (登録日: 2019-08-12, 公開日: 2020-03-18, 最終更新日: 2024-10-23)

主引用文献

Gulen, B.,Rosselin, M.,Fauser, J.,Albers, M.F.,Pett, C.,Krisp, C.,Pogenberg, V.,Schluter, H.,Hedberg, C.,Itzen, A.
Identification of targets of AMPylating Fic enzymes by co-substrate-mediated covalent capture.
Nat.Chem., 12:732-739, 2020

PubMed Abstract: Various pathogenic bacteria use post-translational modifications to manipulate the central components of host cell functions. Many of the enzymes released by these bacteria belong to the large Fic family, which modify targets with nucleotide monophosphates. The lack of a generic method for identifying the cellular targets of Fic family enzymes hinders investigation of their role and the effect of the post-translational modification. Here, we establish an approach that uses reactive co-substrate-linked enzymes for proteome profiling. We combine synthetic thiol-reactive nucleotide derivatives with recombinantly produced Fic enzymes containing strategically placed cysteines in their active sites to yield reactive binary probes for covalent substrate capture. The binary complexes capture their targets from cell lysates and permit subsequent identification. Furthermore, we determined the structures of low-affinity ternary enzyme-nucleotide-substrate complexes by applying a covalent-linking strategy. This approach thus allows target identification of the Fic enzymes from both bacteria and eukarya.

PubMed: 32632184
DOI: 10.1038/s41557-020-0484-6

実験手法

X-RAY DIFFRACTION (2.49 Å)