Monomeric sarcosine oxidase: structure of a covalently flavinylated amine oxidizing enzyme

Trickey, P.; Wagner, Mary Ann; Jörns, Marilyn Schuman; Mathews, F.S.

doi:10.1016/s0969-2126(99)80043-4

Cited by 156 publications

(242 citation statements)

References 60 publications

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“…The region Ala 55 -Asp 60 after helix 2 also shows weak electron density. In MSOX this region corresponds to a flexible loop (Tyr 55 -Tyr 61 ) that changes from a disordered to a weak electron density following the binding of an active site ligand, thus shielding the positive surface potential at the FAD site (12). This loop, and in particular the side chain of Arg 52 , was thus suggested to act in MSOX as a switch for active site accessibility (see below).…”

Section: Resultsmentioning

confidence: 99%

“…3). In pkDAAO O-2 interacts with a threonine and with the partial positive charge of a dipole from helix ␣F5 (10), and in MSOX the flavin O-2 forms a hydrogen bond to the side chain nitrogen of Lys 348 (12).…”

Section: Resultsmentioning

confidence: 99%

“…These interactions serve to stabilize the electrophilic character of the flavin ring, and the negative charge of the anionic form of the semiquinone (1) (12). The proposed candidate is His 45 in MSOX (in GO an alanine is instead present).…”

Section: Resultsmentioning

confidence: 99%

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Structure-Function Correlation in Glycine Oxidase from Bacillus subtilis

Mörtl¹,

Diederichs

Welte

et al. 2004

Journal of Biological Chemistry

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Structure-function relationships of the flavoprotein glycine oxidase (GO), which was recently proposed as the first enzyme in the biosynthesis of thiamine in Bacillus subtilis, has been investigated by a combination of structural and functional studies. The structure of the GO-glycolate complex was determined at 1.8 Å, a resolution at which a sketch of the residues involved in FAD binding and in substrate interaction can be depicted. GO can be considered a member of the "amine oxidase" class of flavoproteins, such as D-amino acid oxidase and monomeric sarcosine oxidase. With the obtained model of GO the monomer-monomer interactions can be analyzed in detail, thus explaining the structural basis of the stable tetrameric oligomerization state of GO, which is unique for the GR 2 subfamily of flavooxidases. On the other hand, the three-dimensional structure of GO and the functional experiments do not provide the functional significance of such an oligomerization state; GO does not show an allosteric behavior. The results do not clarify the metabolic role of this enzyme in B. subtilis; the broad substrate specificity of GO cannot be correlated with the inferred function in thiamine biosynthesis, and the structure does not show how GO could interact with ThiS, the following enzyme in thiamine biosynthesis. However, they do let a general catabolic role of this enzyme on primary or secondary amines to be excluded because the expression of GO is not inducible by glycine, sarcosine, or D-alanine as carbon or nitrogen sources.Glycine oxidase (GO, 1 EC 1.4.3.19) is a flavoprotein consisting of four identical subunits (369 residues each) and containing one molecule of noncovalently bound FAD per 42-kDa protein molecule (1, 2). GO catalyzes a reaction similar to that of D-amino acid oxidase (DAAO, EC 1.4.3.3), a paradigm of the dehydrogenase-oxidase class of flavoproteins (for a recent review see Ref.3). Both enzymes catalyze the oxidative deamination of amino acids to yield the corresponding ␣-imino acids and, after hydrolysis, ␣-keto acids, ammonia (or primary amines), and hydrogen peroxide. Both enzymes show a high pK a for flavin N-3H ionization, do not bind covalently the FAD cofactor, and react readily with sulfite (1-3), but they differ in substrate specificity. In addition to neutral D-amino acids (e.g. D-alanine, D-proline, etc., which are also good substrates of DAAO), GO catalyzes the oxidation of primary and secondary amines (e.g. glycine, sarcosine, etc.) partially sharing the substrate specificity with monomeric sarcosine oxidase (MSOX, EC 1.5.3.1), an enzyme that catalyzes the oxidative demethylation of sarcosine to yield glycine, formaldehyde, and hydrogen peroxide (4). According to investigations of the substrate specificity and of the binding properties, the GO active site seems to preferentially accommodate amines of a small size, such as glycine and sarcosine (1, 2). GO follows a ternary complex sequential mechanism with glycine, sarcosine, and D-proline as substrates in which the rate of product dissociat...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Structure-Function Correlation in Glycine Oxidase from Bacillus subtilis

Mörtl¹,

Diederichs

Welte

et al. 2004

Journal of Biological Chemistry

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“…6 Structures for most of the latter enzymes are available in the PDB, in both the apo form and with various ligands bound. [14][15][16][17] Although the residues of identity vary for each comparison, the sequence identity for the clycine cleavage system T-protein, dimethylglycine oxidase, and sarcosine oxidase is 26, 24, and 22%, respectively. As expected, many of the residues that are conserved are also involved in binding THF (see Fig.…”

Section: Introductionmentioning

confidence: 99%

Structures of dimethylsulfoniopropionate‐dependent demethylase from the marine organism Pelagabacter ubique

et al. 2012

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“…This family is mainly composed of considerably smaller, monomeric proteins (~44 kDa) that contain covalently bound FAD as the only prosthetic group and exhibit modest sequence homology (~20% identity) with the β subunit of TSOX. Members of this family include monomeric sarcosine oxidase (MSOX), N-methyltryptophan oxidase (MTOX), pipecolate oxidase and nikD [11][12][13][14][15][16][17][18] . Crystal structures have been determined for MSOX 11, 14 , a monofunctional enzyme that exhibits sarcosine oxidase but not 5,10-CH 2 -H 4 folate synthase activity 5 .…”

Section: Introductionmentioning

confidence: 99%

Identification of a Stable Flavin-thiolate Adduct in Heterotetrameric Sarcosine Oxidase

Hynson

Mathews

Jörns

2006

Journal of Molecular Biology

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SummaryHeterotetrameric sarcosine oxidase (TSOX) is a complex bifunctional flavoenzyme that contains two flavins. Most of the FMN in recombinant TSOX is present as a covalent adduct with an endogenous ligand. Enzyme denaturation disrupts the adduct, accompanied by release of a stoichiometric amount of sulfide. Enzyme containing ≥ 90% unmodified FMN is prepared by displacement of the endogenous ligand with sulfite, a less tightly bound competing ligand. Reaction of adduct-depleted TSOX with sodium sulfide produces a stable complex that resembled the endogenous TSOX adduct and known 4a-S-cysteinyl flavin adducts. The results provide definitive evidence for sulfide as the endogenous TSOX ligand and strongly suggest that the modified FMN is a 4a-sulfide adduct. A comparable reaction with sodium sulfide is not detected with other flavoprotein oxidases. A model of the postulated TSOX adduct suggests that it is stabilized by nearby residues that may be important in the electron transferase/oxidase function of the coenzyme.

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Monomeric sarcosine oxidase: structure of a covalently flavinylated amine oxidizing enzyme

Cited by 156 publications

References 60 publications

Structure-Function Correlation in Glycine Oxidase from Bacillus subtilis

Structure-Function Correlation in Glycine Oxidase from Bacillus subtilis

Structures of dimethylsulfoniopropionate‐dependent demethylase from the marine organism Pelagabacter ubique

Identification of a Stable Flavin-thiolate Adduct in Heterotetrameric Sarcosine Oxidase

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