Crystal structure of cystathionine β-synthase from honeybee Apis mellifera

Giménez-Mascarell, Paula; Majtán, Tomáš; Oyenarte, Iker; Ereño‐Orbea, June; Majtán, Juraj; Klaudiny, Jaroslav; Kraus, Jan P.; Martínez-Cruz, Luis Alfonso

doi:10.1016/j.jsb.2017.12.008

Cited by 14 publications

(33 citation statements)

References 76 publications

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“…Binding of SAM induces rotation of the two CBS motifs, resulting in reduced interactions with the catalytic core and leading to an open, activated conformation. Strikingly, this conformation is structurally close to those found in the drosophila and honeybee enzymes 3,5 , which are constitutively active, are not soluble upon removal of the C-terminal domain, and do not bind SAM. Yeast CBS, whose three-dimensional structural knowledge is limited to the catalytic core 26 , can bind SAM but its activity is not significantly regulated by the binding of the molecule 9 .…”

supporting

confidence: 54%

“…However, the role of heme in CBS is not fully understood, even if functions in redox sensing and/or enzyme stability and folding have been suggested 10 , 18 – 20 . Notably, the heme-domain has been found only in CBS from higher organisms 1 – 3 , 5 , while CBS enzymes from lower eukaryotic organisms (e.g., S. cerevisiae and T. cruzi 21 ) lack this domain, thus implying that it is not necessary for enzymatic activity (Fig. 1 b).…”

Section: Introductionmentioning

confidence: 99%

“…The reverse transsulfuration pathway is well described in higher organisms [1][2][3][4][5] , where it is responsible for the generation of l-cysteine (l-Cys) from methionine via the intermediates homocysteine (l-Hcys) and cystathionine (l-Cth). The pathway includes two key pyridoxal 5′-phosphate (PLP)-dependent enzymes, namely cystathionine β-synthase (CBS, EC 4.2.1.22) and cystathionine γ-lyase (CGL, EC 4.4.1.1).…”

mentioning

confidence: 99%

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Cystathionine β-synthase is involved in cysteine biosynthesis and H2S generation in Toxoplasma gondii

Conter

Fruncillo

Fernández-Rodríguez

et al. 2020

Sci Rep

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Cystathionine β-synthase (CBS) catalyzes the condensation of serine and homocysteine to water and cystathionine, which is then hydrolyzed to cysteine, α-ketobutyrate and ammonia by cystathionine γ-lyase (CGL) in the reverse transsulfuration pathway. The protozoan parasite Toxoplasma gondii, the causative agent of toxoplasmosis, includes both CBS and CGL enzymes. We have recently reported that the putative T. gondii CGL gene encodes a functional enzyme. Herein, we cloned and biochemically characterized cDNA encoding CBS from T. gondii (TgCBS), which represents a first example of protozoan CBS that does not bind heme but possesses two C-terminal CBS domains. We demonstrated that TgCBS can use both serine and O-acetylserine to produce cystathionine, converting these substrates to an aminoacrylate intermediate as part of a PLP-catalyzed β-replacement reaction. Besides a role in cysteine biosynthesis, TgCBS can also efficiently produce hydrogen sulfide, preferentially via condensation of cysteine and homocysteine. Unlike the human counterpart and similar to CBS enzymes from lower organisms, the TgCBS activity is not stimulated by S-adenosylmethionine. This study establishes the presence of an intact functional reverse transsulfuration pathway in T. gondii and demonstrates the crucial role of TgCBS in biogenesis of H 2 S.

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confidence: 54%

Section: Introductionmentioning

confidence: 99%

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confidence: 99%

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Cystathionine β-synthase is involved in cysteine biosynthesis and H2S generation in Toxoplasma gondii

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Fruncillo

Fernández-Rodríguez

et al. 2020

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“…In addition to these domains, CBSs from higher organisms also have C‐terminal extended regulatory and N‐terminal heme‐binding domains (Finkelstein et al , ; Koracevic and Djordjevic, ; Omura et al , ). In mammals, CBS is activated by the binding of AdoMet to the regulatory domain (Finkelstein et al , ; Koracevic and Djordjevic, ; Prudova et al , ), while CBS proteins from different classes remain constitutively active and do not require AdoMet for the activation (Koutmos et al , ; Gimenez‐Mascarell et al , ). Four CBS structures have been reported, from H. sapiens (Meier et al , ), D. melanogaster (Koutmos et al , ), Apis mellifera (Gimenez‐Mascarell et al , ) and S. cerevisiae (Tu et al , ), and three‐dimentional structural superposition of these CBSs with Hp OCBS showed RMSD values of approximately 2.0 Å for about 300 aligned Cα atoms of the core catalytic domain (i.e., N‐ and C‐terminal domains) (Table and Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The reverse transsulfuration pathway is well characterized in higher organisms such as H. sapiens (Ereno‐Orbea et al , ; Ereno‐Orbea et al , ; McCorvie et al , ), S. cerevisiae (Jhee et al , ), A. mellifera (Gimenez‐Mascarell et al , ) and D. melanogaster (Koutmos et al , ), where it is the sole source of L‐cysteine production from methionine. In bacteria, the reverse transsulfuration pathway is also known as the ‘methionine‐to‐cysteine conversion pathway.’ CBS and CGL have been named, respectively, as MccA and MccB enzymes of the methionine‐to‐cysteine conversion pathway.…”

Section: Discussionmentioning

confidence: 99%

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

Devi

Tarique

Ali

et al. 2019

Molecular Microbiology

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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.

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Current Structural Knowledge on Cystathionine β‐Synthase, a Pivotal Enzyme in the Transsulfuration Pathway

González‐Recio¹,

Fernández-Rodríguez²,

Simón³

et al. 2020

Encyclopedia of Life Sciences

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Cystathionine β‐synthase (CBS), the first enzyme of the reverse transsulfuration pathway, catalyses the pyridoxal‐5′‐phosphate (PLP)‐dependent β‐replacement reaction that condenses l ‐serine with L‐homocysteine to yield cystathionine and water. Besides this canonical reaction and using cysteine and homocysteine as substrates, CBS can also efficiently produce hydrogen sulfide (H 2 S) through alternative β‐replacement and β‐elimination processes. The structural information on the full‐length enzyme has remained elusive for decades and is still very scarce, but some advances in the recent years have uncovered its peculiar modular architecture, provided a glimpse of its conformational landscape and revealed some of the reaction intermediates formed during the catalysis. All these data have helped comprehend, at least partially, the regulatory mechanisms and the catalytic abilities of the enzyme across different organisms. This article aims to overview the current information on the CBS structure from its most sophisticated variants found in mammals to its simplest homologs in bacteria. A more detailed understanding of CBS structure and function is needed, which could subsequently serve as a basis for the development of drugs to treat human diseases, such as CBS‐deficient homocystinuria, Alzheimer diseases and some cancers, as well as of new antibiotics against multidrug‐resistant pathogenic bacteria. Key Concepts Reverse transsulfuration is a two‐step metabolic route that allows the conversion of the essential amino acid methionine into cysteine. This process generates hydrogen sulfide and impedes the accumulation of the toxic intermediate homocysteine. Hydrogen sulfide is an important gasotransmitter involved in physiological functions such as neuroprotection and the regulation of blood pressure. Homocystinuria consists of the abnormal accumulation of homocysteine, and is an inherited disorder due to the deficient activity of CBS. This pathology causes vascular thromboses, skeletal defects, mental retardation, and even early death. The three‐dimensional structure of CBS provides a suitable template to develop drugs to treat homocystinuria and related pathologies in humans. The comparative structural analysis of the CBS enzymes from different organisms are key to intervene in their sulfur metabolism, and thus represents a potential therapeutic approach against pathogens.

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Crystal structure of cystathionine β-synthase from honeybee Apis mellifera

Cited by 14 publications

References 76 publications

Cystathionine β-synthase is involved in cysteine biosynthesis and H2S generation in Toxoplasma gondii

Cystathionine β-synthase is involved in cysteine biosynthesis and H2S generation in Toxoplasma gondii

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

Current Structural Knowledge on Cystathionine β‐Synthase, a Pivotal Enzyme in the Transsulfuration Pathway

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