The Transition-State Structure for Human MAT2A from Isotope Effects

Firestone, Ross S.; Schramm, Vern L.

doi:10.1021/jacs.7b05803

Cited by 19 publications

(40 citation statements)

References 39 publications

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“…C) indicates that the early step of reaction has taken place, but the S114A mutant fails to provide the nucleophilic attack of methionine's sulfur atom against C'5 atom of AMP‐PNP, thus unable to produce SAMe. These observations are consistent with the suggestion that C‐O bond breaking takes place prior to C‐S bond formation at the catalytic transition state of human wt MATα2 . Previously our high‐resolution structure of SAMe+ADO+MET+PPNP wt MATα2 (PDB: http://www.rcsb.org/pdb/search/structidSearch.do?structureId=5A1I) had revealed that the Ser114 is the sole residue in the gating loop that interacts with AMP‐PNP via a water molecule.…”

Section: Resultssupporting

confidence: 88%

Control and regulation of S‐Adenosylmethionine biosynthesis by the regulatory β subunit and quinolone‐based compounds

et al. 2019

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Methylation is an underpinning process of life and provides control for biological processes such as DNA synthesis, cell growth, and apoptosis. Methionine adenosyltransferases (MAT) produce the cellular methyl donor, S‐Adenosylmethionine (SAMe). Dysregulation of SAMe level is a relevant event in many diseases, including cancers such as hepatocellular carcinoma and colon cancer. In addition, mutation of Arg264 in MATα1 causes isolated persistent hypermethioninemia, which is characterized by low activity of the enzyme in liver and high level of plasma methionine. In mammals, MATα1/α2 and MATβV1/V2 are the catalytic and the major form of regulatory subunits, respectively. A gating loop comprising residues 113–131 is located beside the active site of catalytic subunits (MATα1/α2) and provides controlled access to the active site. Here, we provide evidence of how the gating loop facilitates the catalysis and define some of the key elements that control the catalytic efficiency. Mutation of several residues of MATα2 including Gln113, Ser114, and Arg264 lead to partial or total loss of enzymatic activity, demonstrating their critical role in catalysis. The enzymatic activity of the mutated enzymes is restored to varying degrees upon complex formation with MATβV1 or MATβV2, endorsing its role as an allosteric regulator of MATα2 in response to the levels of methionine or SAMe. Finally, the protein–protein interacting surface formed in MATα2:MATβ complexes is explored to demonstrate that several quinolone‐based compounds modulate the activity of MATα2 and its mutants, providing a rational for chemical design/intervention responsive to the level of SAMe in the cellular environment.EnzymesMethionine adenosyltransferase (http://www.chem.qmul.ac.uk/iubmb/enzyme/EC2/5/1/6.html).DatabaseStructural data are available in the RCSB PDB database under the PDB ID http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6FBN (Q113A), http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6FBP (S114A: P22121), http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6FBO (S114A: I222), http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6FCB (P115G), http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6FCD (R264A), http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6FAJ (wtMATα2: apo), http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6G6R (wtMATα2: holo)

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Section: Resultssupporting

confidence: 88%

Control and regulation of S‐Adenosylmethionine biosynthesis by the regulatory β subunit and quinolone‐based compounds

et al. 2019

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“…A solvent accessible active site of MAT enables this enzyme to tolerate analogues of methionine and (seleno)methionine substrates bearing longer alkyl chains and functional handles on S/Se-centres producing a suite of analogues such as 7-9 ( Figure 2). [21,25,39,[47][48][49] The in situ synthesis of these cofactor analogues can then be used as substrates to transfer functional handles catalyzed by a variety of O- (14 a-d), C-(14 e) and N-(14 g) MTases (Figure 4a). [9,[20][21]25,[37][38][39] In addition, Seebeck et al expanded their two-enzyme SAM regeneration strategy (Figure 3B) [35] to stereoselectively prepare L-or D-β-alkylated αamino acids amino acids such as 14 f ( Figure 4B), [40] which are challenging to prepare by conventional chemical synthesis.…”

Section: Enhancing the Scope Of Biocatalytic Alkylation Using Modifiementioning

confidence: 99%

“…An atom-efficient alternative to the use of ATP as the adenosyl donor is to use 5'-chloro-5'-deoxyadenosine (ClDA) catalyzed by the chlorinase SalL (Salinispora tropica, Figure 4C). [50] Although the substrate scope of wildtype SalL is narrower than of MAT, [20][21]37,39,44,[47][48] SalL is capable of SAM analogue synthesis and is compatible with a range of small molecule (14 h,j,k) [23][24]27] and DNA (14 i) [22,24] MTases, thereby enhancing the flexibility of enzymatic alkylation routes available ( Figure 4A, B).…”

Section: Enhancing the Scope Of Biocatalytic Alkylation Using Modifiementioning

confidence: 99%

“…In nature, MAT uses ATP as the adenosyl donor and methionine to prepare SAM. A solvent accessible active site of MAT enables this enzyme to tolerate analogues of methionine and (seleno)methionine substrates bearing longer alkyl chains and functional handles on S / Se ‐centres producing a suite of analogues such as 7 – 9 (Figure ) . The in situ synthesis of these cofactor analogues can then be used as substrates to transfer functional handles catalyzed by a variety of O‐ ( 14 a – d ), C‐ ( 14 e ) and N‐ ( 14 g ) MTases (Figure a) .…”

Section: Enhancing the Scope Of Biocatalytic Alkylation Using Modifiementioning

confidence: 99%

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Biocatalytic Alkylation Cascades: Recent Advances and Future Opportunities for Late‐Stage Functionalization

2020

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This Concept article describes the latest developments in the emerging area of late-stage biocatalytic alkylation. Central to these developments is the ability to efficiently prepare Sadenosyl methionine (SAM) cofactor analogues and couple this with enzymatic alkyl transfer. Recent developments in the enzymatic synthesis of SAM cofactor analogues are summarized first, followed by their application as alkyl transfer agents catalyzed by methyltransferases (MTases). Second, innovative methods to regenerate SAM cofactors by enzymatic cascades is reported. Finally, future opportunities towards establishing a generalized platform for late-stage alkylation are described.

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“…MATV1 was able to alter the activity of MAT2 without changing its catalytic transition state (Firestone & Schramm, 2017;Murray et al, 2014), and the catalytic site of MAT2 was also preserved in the presence of MAT (Murray et al, 2016), indicating that the increase in activity is modulated by allosteric regulation. The restoration of the activity of the MAT1 R264H mutant by the regulatory subunit MATV1 or the compound SCR0911 suggests a similar binding interface.…”

Section: The R264h Mutation Weakens the Dimer-dimer Interface In Matamentioning

confidence: 99%

Structural basis of the dominant inheritance of hypermethioninemia associated with the Arg264His mutation in the MAT1A gene

Panmanee¹,

Antonyuk²,

Hasnain³

2020

Acta Crystallogr D Struct Biol

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Methionine adenosyltransferase (MAT) deficiency, characterized by isolated persistent hypermethioninemia (IPH), is caused by mutations in the MAT1A gene encoding MATl, one of the major hepatic enzymes. Most of the associated hypermethioninemic conditions are inherited as autosomal recessive traits; however, dominant inheritance of hypermethioninemia is caused by an Arg264His (R264H) mutation. This mutation has been confirmed in a screening programme of newborns as the most common mutation in babies with IPH. Arg264 makes an inter-subunit salt bridge located at the dimer interface where the active site assembles. Here, it is demonstrated that the R264H mutation results in greatly reduced MAT activity, while retaining its ability to dimerize, indicating that the lower activity arises from alteration at the active site. The first crystallographic structure of the apo form of the wild-type MATl enzyme is provided, which shows a tetrameric assembly in which two compact dimers combine to form a catalytic tetramer. In contrast, the crystal structure of the MATl R264H mutant reveals a weaker dimeric assembly, suggesting that the mutation lowers the affinity for dimer-dimer interaction. The formation of a hetero-oligomer with the regulatory MATV1 subunit or incubation with a quinolone-based compound (SCR0911) results in the near-full recovery of the enzymatic activity of the pathogenic mutation R264H, opening a clear avenue for a therapeutic solution based on chemical interventions that help to correct the defect of the enzyme in its ability to metabolize methionine. research papers Acta Cryst. (2020). D76, 594-607 Panmanee et al. Structural basis of MAT1-linked hypermethioninemia 595

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The Transition-State Structure for Human MAT2A from Isotope Effects

Cited by 19 publications

References 39 publications

Control and regulation of S‐Adenosylmethionine biosynthesis by the regulatory β subunit and quinolone‐based compounds

Control and regulation of S‐Adenosylmethionine biosynthesis by the regulatory β subunit and quinolone‐based compounds

Biocatalytic Alkylation Cascades: Recent Advances and Future Opportunities for Late‐Stage Functionalization

Structural basis of the dominant inheritance of hypermethioninemia associated with the Arg264His mutation in the MAT1A gene

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