The PAPS-Independent Aryl Sulfotransferase and the Alternative Disulfide Bond Formation System in Pathogenic Bacteria

Abstract

Sulfurylation of biomolecules (often termed sulfonation or sulfation) has been described in many organisms in all kingdoms of life. To date, most studies on sulfotransferases, the enzymes catalyzing sulfurylation, have focused on 3'-phosphate-5'-phosphosulfate (PAPS)-dependent enzymes, which transfer the sulfuryl group from this activated anhydride to hydroxyl groups of acceptor molecules. By contrast, the PAPS-independent aryl sulfotransferases (ASSTs) from bacteria, which catalyze sulfotransfer from phenolic sulfate esters to another phenol in the bacterial periplasm, were not well characterized until recently, although they were first described in 1986 in a search for nonhepatic sulfurylation processes. Recent studies revealed that this unusual class of sulfotransferases differs profoundly in both molecular structure and catalytic mechanism from PAPS-dependent sulfotransferases, and that ASSTs from certain bacterial pathogens are upregulated during infection. In this review, we summarize the literature on the roles of sulfurylation in prokaryotes and analyze the occurrence of ASSTs and their dependence on Dsb proteins catalyzing oxidative folding in the periplasm. Furthermore, we discuss structural differences and similarities between aryl sulfotransferases and PAPS-dependent sulfotransferases.

High-throughput cell-based assays for the preclinical development of DsbA inhibitors as antivirulence therapeutics

“…AssT is a large periplasmic enzyme encoded by UPEC and other intestinal bacteria (29) that was reported to be upregulated in the urine of UPEC-infected mice (46, 47), but was not required for colonisation of the murine bladder (29). The gene encoding AssT is found in a tri-cistronic operon with the dsbL and dsbI genes, which encode an accessory redox protein pair in UPEC with specificity for AssT (39, 48), although the DsbA and DsbB redox pair was also shown to functionally fold AssT (27, 29). The AssT activity assay was previously performed in liquid medium using bacterial cell lysates (39) or on solid medium using whole live cells (27).…”

A robust protocol for directed aryl sulfotransferase evolution toward the carbohydrate building block GlcNAc

“…Two classes of sulfotransferases can be distinguished according to their specificity toward the donor molecule, 3′‐phosphoadenosine‐5′‐phosphosulfate (PAPS)‐dependent and PAPS‐independent sulfotransferases (Malojcić & Glockshuber, ). PAPS‐dependent sulfotransferases are reported, in contrast to PAPS‐independent sulfotransferases, to regioselectively sulfurylate proteins, lipids, and polysaccharides (see excellent review by Wong et al, ).…”

“…An attractive alternative are PAPS‐independent sulfotransferases such as prokaryotic aryl sulfotransferases (AST; also known as aryl sulfate sulfotransferases; EC 2.8.2.22). ASTs use phenolic sulfate esters (e.g., p ‐nitrophenyl sulfate ( p NPS), 4‐methylumbelliferyl sulfate (4‐MUS) and 1‐naphthyl sulfate) as sulfuryl donors (Kobashi, Kim, & Morikawa, ; Malojcić & Glockshuber, ) that are more cost‐efficient than PAPS. Prokaryotic sulfotransferase have usually a broad substrate scope (Kim, Yoon, Koizumi, & Kobashi, ; van der Horst et al, ) and bacterial expression systems can be used for enzyme production (Kim, Hyun, Lee, Kobashi, & Kim, ) and reengineering (Koryakina, Neville, Nonaka, Van Lanen, & Williams, ).…”

Substrate Engineering and its Synthetic Utility in the Sulfation of Primary Aliphatic Alcohol Groups by a Bacterial Arylsulfotransferase

“…[6,7,8] Another class of enzymes that also catalyze sulfation reactions are the arylsulfotransferases (ASTs) from bacterial origin.T hese enzymes use sulfated phenols as donor, e.g., p-nitrophenyl sulfate (p-NPS) and have broad substrate specificity thoughi ng eneral only phenolic groups are sulfated. [10][11][12][13] We have usedt he recombinant enzyme from Desulfitobacterium hafniense in the synthesis of av ariety of sulfatedc ompounds,e .g.,e strone, estradiol,e nkephalin and polyphenols. [14,15] We also discovered and in great contrast to all other known bacterial ASTs,t hat this enzyme was able to use av ariety of non-phenolic alcohols as sulfate acceptor although the conversions were low.…”

“…[21,22] Thesee nzymes catalyze transglycosylation reactions and this paves the wayt ot he formation of more complexs ulfatedc arbohydrates.T his aromatic nitro group also allowsamore convenient isolation and purificationo ft he sulfated compounds and analysis by HPLC is more easy.U sing the above procedure we were able to synthesize as ulfated glycerol derivative andt hree sulfated carbohydrate derivatives (Scheme 2) which were characterized by 13 Ca nd 1 HNMR (see the Supporting Information).…”