Purification and properties of cobalamin-independent methionine synthase from Candida albicans and Saccharomyces cerevisiae

Suliman, Huda S.; Sawyer, Gregory M.; Appling, Dean R.; Robertus, Jon D.

doi:10.1016/j.abb.2005.06.016

Cited by 26 publications

(36 citation statements)

References 28 publications

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“…Mammals utilize the cobalamin-dependent methionine synthase, while plants and fungi draw on the cobalamin-independent enzyme. Therefore, having demonstrated that the methionine synthase is essential for A. fumigatus pathogenicity, we propose that it constitutes a specific and promising fungal target for chemotherapy, as has been suggested previously (79,96), which would actually have a broad antifungal spectrum.…”

Section: Discussionmentioning

confidence: 99%

Exploration of Sulfur Assimilation of Aspergillus fumigatus Reveals Biosynthesis of Sulfur-Containing Amino Acids as a Virulence Determinant

et al. 2016

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e Fungal infections are of major relevance due to the increased numbers of immunocompromised patients, frequently delayed diagnosis, and limited therapeutics. To date, the growth and nutritional requirements of fungi during infection, which are relevant for invasion of the host, are poorly understood. This is particularly true for invasive pulmonary aspergillosis, as so far, sources of (macro)elements that are exploited during infection have been identified to only a limited extent. Here, we have investigated sulfur (S) utilization by the human-pathogenic mold Aspergillus fumigatus during invasive growth. Our data reveal that inorganic S compounds or taurine is unlikely to serve as an S source during invasive pulmonary aspergillosis since a sulfate transporter mutant strain and a sulfite reductase mutant strain are fully virulent. In contrast, the S-containing amino acid cysteine is limiting for fungal growth, as proven by the reduced virulence of a cysteine auxotroph. Moreover, phenotypic characterization of this strain further revealed the robustness of the subordinate glutathione redox system. Interestingly, we demonstrate that methionine synthase is essential for A. fumigatus virulence, defining the biosynthetic route of this proteinogenic amino acid as a potential antifungal target. In conclusion, we provide novel insights into the nutritional requirements of A. fumigatus during pathogenesis, a prerequisite to understanding and fighting infection.

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Section: Discussionmentioning

confidence: 99%

Exploration of Sulfur Assimilation of Aspergillus fumigatus Reveals Biosynthesis of Sulfur-Containing Amino Acids as a Virulence Determinant

et al. 2016

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“…The yield of CH 2 -THF was determined by solving the equilibria of THF, formaldehyde, and ␤-mercaptoethanol (15). The preparation of CH ϩ -THF from 5-CHO-THF (MP Biomedicals, Solon, OH) was conducted as described previously (16 (17). Further preparation of 5,10-CH 2 -H 4 PteGlu 5 was accomplished by incubation with formaldehyde as described previously (14).…”

Section: Methodsmentioning

confidence: 99%

“…Following PCR, rMTHFD2L-x8 was subcloned into YEp24ES (11,17) using the HindIII and BamHI restriction sites. Plasmids were transformed into the yeast strain MWY4.5 (ser1 ura3-52 trp1 leu2 his4 ade3-30/65 ⌬mtd1), and transformants were selected for uracil prototrophy.…”

Section: Methodsmentioning

confidence: 99%

“…The resulting PCR-amplified fragment was cloned into the pET22b vector (Novagen, EMD Biosciences). The pET22b-rMTHFD2L⌬1-40 was further subcloned into a YEp24-based yeast expression vector containing the Saccharomyces cerevisiae MET6 promoter using sequence-and ligation-independent cloning (17,19). The YEp24 vector was modified to include a multiple cloning site and an N-terminal His 6 tag (11).…”

Section: Methodsmentioning

confidence: 99%

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Mitochondrial MTHFD2L Is a Dual Redox Cofactor-specific Methylenetetrahydrofolate Dehydrogenase/Methenyltetrahydrofolate Cyclohydrolase Expressed in Both Adult and Embryonic Tissues

Shin

Bryant

Momb

et al. 2014

Journal of Biological Chemistry

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“…Expression in Yeast and Enzyme Assay-The full-length rat cDNA was subcloned from pET22b-rMTHFD2L using sequence-and ligation-independent cloning (34) into a YEp24-based yeast expression vector containing the Saccharomyces cerevisiae MET6 promoter (42). This vector (Yep24ES) was modified by Everett Stone (University of Texas, Austin, TX) to include a multiple cloning site and an N-terminal His 6 tag.…”

Section: Alkaline Carbonate Extraction and Protease Treatment Ofmentioning

confidence: 99%

Mammalian MTHFD2L Encodes a Mitochondrial Methylenetetrahydrofolate Dehydrogenase Isozyme Expressed in Adult Tissues

Swetha

Young²,

Cole

et al. 2011

Journal of Biological Chemistry

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Previous studies in our laboratory showed that isolated, intact adult rat liver mitochondria are able to oxidize the 3-carbon of serine and the N-methyl carbon of sarcosine to formate without the addition of any other cofactors or substrates. Conversion of these 1-carbon units to formate requires several folate-interconverting enzymes in mitochondria. The enzyme(s) responsible for conversion of 5,10-methylene-tetrahydrofolate (CH 2 -THF) to 10-formyl-THF in adult mammalian mitochondria are currently unknown. A new mitochondrial CH 2 -THF dehydrogenase isozyme, encoded by the MTHFD2L gene, has now been identified. The recombinant protein exhibits robust NADP ؉ -dependent CH 2 -THF dehydrogenase activity when expressed in yeast. The enzyme is localized to mitochondria when expressed in CHO cells and behaves as a peripheral membrane protein, tightly associated with the matrix side of the mitochondrial inner membrane. The MTHFD2L gene is subject to alternative splicing and is expressed in adult tissues in humans and rodents. This CH 2 -THF dehydrogenase isozyme thus fills the remaining gap in the pathway from CH 2 -THF to formate in adult mammalian mitochondria.

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Purification and properties of cobalamin-independent methionine synthase from Candida albicans and Saccharomyces cerevisiae

Cited by 26 publications

References 28 publications

Exploration of Sulfur Assimilation of Aspergillus fumigatus Reveals Biosynthesis of Sulfur-Containing Amino Acids as a Virulence Determinant

Exploration of Sulfur Assimilation of Aspergillus fumigatus Reveals Biosynthesis of Sulfur-Containing Amino Acids as a Virulence Determinant

Mitochondrial MTHFD2L Is a Dual Redox Cofactor-specific Methylenetetrahydrofolate Dehydrogenase/Methenyltetrahydrofolate Cyclohydrolase Expressed in Both Adult and Embryonic Tissues

Mammalian MTHFD2L Encodes a Mitochondrial Methylenetetrahydrofolate Dehydrogenase Isozyme Expressed in Adult Tissues

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