Investigation of the enzymes required for the biosynthesis of an unusual formylated sugar in the emerging human pathogen <scp><i>Helicobacter canadensis</i></scp>

Heisdorf, C.J.; Griffiths, W.A.D.; Thoden, James B.; Holden, Hazel M.

doi:10.1002/pro.4169

Cited by 4 publications

(4 citation statements)

References 58 publications

(135 reference statements)

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“…Thus, we postulated that a probable substrate for the H. pullorum N ‐acetyltransferase was either dTDP‐3‐amino‐3,6‐dideoxy‐ d ‐glucose or dTDP‐3‐amino‐3,6‐dideoxy‐ d ‐galactose. In a previous investigation, we demonstrated that the 3,4‐ketoisomerase from H. canadensis produces dTDP‐3‐keto‐3,6‐dideoxy‐ d ‐glucose 21 . The amino acid sequence similarities and identities between the H. canadensis and the H. pullorum 3,4‐ketoisomerases are 44% and 66%, respectively.…”

Section: Resultsmentioning

confidence: 94%

“…In a previous investigation, we demonstrated that the 3,4-ketoisomerase from H. canadensis produces dTDP-3-keto-3,6-dideoxy-D-glucose. 21 The amino acid sequence similarities and identities between the H. canadensis and the H. pullorum 3,4-ketoisomerases are 44% and 66%, respectively.…”

Section: Kinetic Properties Of the H Pullorum N-acetyltransferasementioning

confidence: 99%

“…Kinetic parameters are listed in Table 2. 3.9 | Production of dTDP-3-amino-3,6-dideoxy-D-glucose and dTDP-3-amino-3,6-dideoxy-D-galactose These ligands were synthesized and purified as previously reported from this laboratory. 21…”

Section: Determination Of Kinetic Parametersmentioning

confidence: 99%

“…20 A recent investigation in our laboratory demonstrated that the genome of Helicobacter canadensis encodes for proteins required for the biosynthesis of N-formylated sugars. 21 With that in mind, we searched the same genome for possible genes encoding N-acetyltransferases, and indeed, we located one. All attempts to produce purified protein for a subsequent biochemical analysis were unsuccessful.…”

Section: Introductionmentioning

confidence: 99%

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Biochemical investigation of an N‐acetyltransferase from Helicobacter pullorum

et al. 2021

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N-acetylated sugars are often found, for example, on the lipopolysaccharides of Gram-negative bacteria, on the S-layers of Gram-positive bacteria, and on the capsular polysaccharides. Key enzymes involved in their biosynthesis are the sugar N-acetyltransferases. Here, we describe a structural and functional analysis of one such enzyme from Helicobacter pullorum, an emerging pathogen that may be associated with gastroenteritis and gallbladder and liver diseases. For this analysis, the gene BA919-RS02330 putatively encoding an N-acetyltransferase was cloned, and the corresponding protein was expressed and purified. A kinetic analysis demonstrated that the enzyme utilizes dTDP-3-amino-3,6-dideoxy-D-glucose as a substrate as well as dTDP-3-amino-3,6-dideoxy-D-galactose, albeit at a reduced rate. In addition to this kinetic analysis, a similar enzyme from Helicobacter bilis was cloned and expressed, and its kinetic parameters were determined. Seven X-ray crystallographic structures of various complexes of the H. pullorum wild-type enzyme (or the C80T variant) were determined to resolutions of 1.7 Å or higher. The overall molecular architecture of the H. pullorum N-acetyltransferase places it into the Class II left-handedβ-helix superfamily (LβH). Taken together, the data presented herein suggest that 3-acetamido-3,6-dideoxy-D-glucose (or the galactose derivative) is found on either the H. pullorum O-antigen or in another of its complex glycoconjugates. A BLAST search suggests that more than 50 non-pylori Helicobacter spp. have genes encoding N-acetyltransferases. Given that there is little information concerning the complex glycans in non-pylori Helicobacter spp. and considering their zoonotic potential, our results provide new biochemical insight into these pathogens

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Section: Resultsmentioning

confidence: 94%

Section: Kinetic Properties Of the H Pullorum N-acetyltransferasementioning

confidence: 99%

Section: Determination Of Kinetic Parametersmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Biochemical investigation of an N‐acetyltransferase from Helicobacter pullorum

et al. 2021

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Investigation of the enzymes required for the biosynthesis of an unusual formylated sugar in the emerging human pathogen Helicobacter canadensis

et al. 2021

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It is now well established that the Gram-negative bacterium, Helicobacter pylori, causes gastritis in humans. In recent years, it has become apparent that the so-called non-pylori Helicobacters, normally infecting pigs, cats, and dogs, may also be involved in human pathology via zoonotic transmission. Indeed, more than 30 species of non-pylori Helicobacters have been identified thus far. One such organism is Helicobacter canadensis, an emerging pathogen whose

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