3ETS

Crystal structure of a bacterial arylsulfate sulfotransferase catalytic intermediate with 4-methylumbelliferone bound in the active site

Summary for 3ETS

• Entry DOI: 10.2210/pdb3ets/pdb
• Related: 3ELQ 3ETT
• Descriptor: Arylsulfate sulfotransferase, 7-hydroxy-4-methyl-2H-chromen-2-one, SULFATE ION, ... (4 entities in total)
• Functional Keywords: beta propeller, sulfohistidine, protein-substrate complex, periplasm, transesterification, sulfate, phenol, bacteria, transferase 4-methylumbelliferone, 4-methylumbelliferylsulfate, transferase
• Biological source: Escherichia coli
• Total number of polymer chains: 2
• Total formula weight: 128782.12

Authors

Malojcic, G.,Owen, R.L.,Grimshaw, J.P.,Glockshuber, R. (deposition date: 2008-10-08, release date: 2008-11-25, Last modification date: 2023-09-06)

Primary citation

Malojcic, G.,Owen, R.L.,Grimshaw, J.P.,Brozzo, M.S.,Dreher-Teo, H.,Glockshuber, R.
A structural and biochemical basis for PAPS-independent sulfuryl transfer by aryl sulfotransferase from uropathogenic Escherichia coli.
Proc.Natl.Acad.Sci.USA, 105:19217-19222, 2008

PubMed Abstract: Sulfotransferases are a versatile class of enzymes involved in numerous physiological processes. In mammals, adenosine 3'-phosphate-5'-phosphosulfate (PAPS) is the universal sulfuryl donor, and PAPS-dependent sulfurylation of small molecules, including hormones, sugars, and antibiotics, is a critical step in hepatic detoxification and extracellular signaling. In contrast, little is known about sulfotransferases in bacteria, which make use of sulfurylated molecules as mediators of cell-cell interactions and host-pathogen interactions. Bacterial arylsulfate sulfotransferases (also termed aryl sulfotransferases), in contrast to PAPS-dependent sulfotransferases, transfer sulfuryl groups exclusively among phenolic compounds in a PAPS-independent manner. Here, we report the crystal structure of the virulence factor arylsulfate sulfotransferase (ASST) from the prototypic, pyelonephritogenic Escherichia coli strain CFT073 at 2.0-A resolution, and 2 catalytic intermediates, at 2.1-A and 2.4-A resolution, with substrates bound in the active site. ASST is one of the largest periplasmic enzymes and its 3D structure differs fundamentally from all other structurally characterized sulfotransferases. Each 63.8-kDa subunit of the ASST homodimer comprises a 6-bladed beta-propeller domain and a C-terminal beta-sandwich domain. The active sites of the dimer are situated at the center of the channel formed by each beta-propeller and are defined by the side chains of His-252, His-356, Arg-374, and His-436. We show that ASST follows a ping-pong bi-bi reaction mechanism, in which the catalytic residue His-436 undergoes transient sulfurylation, a previously unreported covalent protein modification. The data provide a framework for understanding PAPS-independent sulfotransfer and a basis for drug design targeting this bacterial virulence factor.

PubMed: 19036922
DOI: 10.1073/pnas.0806997105

Experimental method

X-RAY DIFFRACTION (2.4 Å)