4P07

Bacterial aryl sulfotransferase (ASST) soaked with human urine

4P07 の概要

• エントリーDOI: 10.2210/pdb4p07/pdb
• 関連するPDBエントリー: 3ELQ 3ETS 3ETT 4P04 4P05 4P06
• 分子名称: Arylsulfate sulfotransferase AssT, SULFATE ION (3 entities in total)
• 機能のキーワード: sulfotransferase, beta propeller, active site mutant, transferase
• 由来する生物種: Escherichia coli CFT073
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 127885.41

構造登録者

Malojcic, G.,Owen, R.L.,Glockshuber, R. (登録日: 2014-02-20, 公開日: 2014-03-26, 最終更新日: 2024-10-23)

主引用文献

Malojcic, G.,Owen, R.L.,Glockshuber, R.
Structural and mechanistic insights into the PAPS-independent sulfotransfer catalyzed by bacterial aryl sulfotransferase and the role of the DsbL/Dsbl system in its folding.
Biochemistry, 53:1870-1877, 2014

PubMed Abstract: Bacterial aryl sulfotransferases (ASSTs) catalyze sulfotransfer from a phenolic sulfate to a phenol. These enzymes are frequently found in pathogens and upregulated during infection. Their mechanistic understanding is very limited, and their natural substrates are unknown. Here, the crystal structures of Escherichia coli CFT073 ASST trapped in its presulfurylation state with model donor substrates bound in the active site are reported, which reveal the molecular interactions governing substrate recognition. Furthermore, spectroscopic titrations with donor substrates and sulfurylation kinetics of ASST illustrate that this enzyme binds substrates in a 1:1 stoichiometry and that the active sites of the ASST homooligomer act independently. Mass spectrometry and crystallographic experiments of ASST incubated with human urine demonstrate that urine contains a sulfuryl donor substrate. In addition, we examined the capability of the two paralogous dithiol oxidases present in uropathogenic E. coli CFT073, DsbA, and the ASST-specific enzyme DsbL, to introduce the single, conserved disulfide bond into ASST. We show that DsbA and DsbL introduce the disulfide bond into unfolded ASST at similar rates. Hence, a chaperone effect of DsbL, not present in DsbA, appears to be responsible for the dependence of efficient ASST folding on DsbL in vivo. The conservation of paralogous dithiol oxidases with different substrate specificities in certain bacterial strains may therefore be a consequence of the complex folding pathways of their substrate proteins.

PubMed: 24601529
DOI: 10.1021/bi401725j

実験手法

X-RAY DIFFRACTION (2.59 Å)