Identification and characterisation of Helicobacter pylori O-acetylserine-dependent cystathionine β-synthase, a distinct member of the PLP-II family

Suneeta Devi, Khaja Faisal Tarique, Mohammad Farhan Ali, Syed Arif Abdul Rehman, Samudrala Gourinath (Lead / Corresponding author)

Research output: Contribution to journalArticle

Abstract

O-acetylserine sulfhydrylase (OASS) and cystathionine β-synthase (CBS) are members of the PLP-II family, and involved in L-cysteine production. OASS produces L-cysteine via a de novo pathway while CBS participates in the reverse transsulfuration pathway. O-acetylserine-dependent CBS (OCBS) was previously identified as a new member of the PLP-II family, which are predominantly seen in bacteria. The bacterium Helicobacter pylori possess only one OASS (hp0107) gene and we showed that the protein coded by this gene actually functions as an OCBS and utilizes L-homocysteine and O-acetylserine (OAS) to produce cystathionine. HpOCBS did not show CBS activity with the substrate L-serine and required OAS exclusively. The HpOCBS structure in complex with methionine showed a closed cleft state, explaining the initial mode of substrate binding. Sequence and structural analyses showed differences between the active sites of OCBS and CBS, and explain their different substrate preferences. We identified three hydrophobic residues near the active site of OCBS, corresponding to one serine and two tyrosine residues in CBSs. Mutational studies were performed on HpOCBS and Saccharomyces cerevisiae CBS. A ScCBS double mutant (Y158F/Y226V) did not display activity with L-serine, indicating indispensability of these polar residues for selecting substrate L-serine, however, did show activity with OAS.

Original languageEnglish
Pages (from-to)718-739
Number of pages22
JournalMolecular Microbiology
Volume112
Issue number2
Early online date27 May 2019
DOIs
Publication statusPublished - 21 Aug 2019

Fingerprint

Cystathionine
Helicobacter pylori
Cysteine Synthase
Serine
Cysteine
Catalytic Domain
Bacteria
Homocysteine
O-acetylserine
Methionine
Sequence Analysis
Saccharomyces cerevisiae
Tyrosine

Keywords

  • Cystathionine β‐synthase
  • Helicobacter pylori
  • Hydrogen sulfide
  • O‐acetylserine sulfhydrylase
  • pyridoxal 5’‐phosphate
  • reverse transsulfuration pathway

Cite this

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title = "Identification and characterisation of Helicobacter pylori O-acetylserine-dependent cystathionine β-synthase, a distinct member of the PLP-II family",
abstract = "O-acetylserine sulfhydrylase (OASS) and cystathionine β-synthase (CBS) are members of the PLP-II family, and involved in L-cysteine production. OASS produces L-cysteine via a de novo pathway while CBS participates in the reverse transsulfuration pathway. O-acetylserine-dependent CBS (OCBS) was previously identified as a new member of the PLP-II family, which are predominantly seen in bacteria. The bacterium Helicobacter pylori possess only one OASS (hp0107) gene and we showed that the protein coded by this gene actually functions as an OCBS and utilizes L-homocysteine and O-acetylserine (OAS) to produce cystathionine. HpOCBS did not show CBS activity with the substrate L-serine and required OAS exclusively. The HpOCBS structure in complex with methionine showed a closed cleft state, explaining the initial mode of substrate binding. Sequence and structural analyses showed differences between the active sites of OCBS and CBS, and explain their different substrate preferences. We identified three hydrophobic residues near the active site of OCBS, corresponding to one serine and two tyrosine residues in CBSs. Mutational studies were performed on HpOCBS and Saccharomyces cerevisiae CBS. A ScCBS double mutant (Y158F/Y226V) did not display activity with L-serine, indicating indispensability of these polar residues for selecting substrate L-serine, however, did show activity with OAS.",
keywords = "Cystathionine β‐synthase, Helicobacter pylori, Hydrogen sulfide, O‐acetylserine sulfhydrylase, pyridoxal 5’‐phosphate, reverse transsulfuration pathway",
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Identification and characterisation of Helicobacter pylori O-acetylserine-dependent cystathionine β-synthase, a distinct member of the PLP-II family. / Devi, Suneeta; Tarique, Khaja Faisal; Ali, Mohammad Farhan; Abdul Rehman, Syed Arif ; Gourinath, Samudrala (Lead / Corresponding author).

In: Molecular Microbiology, Vol. 112, No. 2, 21.08.2019, p. 718-739.

Research output: Contribution to journalArticle

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T1 - Identification and characterisation of Helicobacter pylori O-acetylserine-dependent cystathionine β-synthase, a distinct member of the PLP-II family

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AU - Tarique, Khaja Faisal

AU - Ali, Mohammad Farhan

AU - Abdul Rehman, Syed Arif

AU - Gourinath, Samudrala

N1 - © 2019 John Wiley & Sons Ltd.

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