Integrated metabolomics and proteomics analysis to provide insights into muscle atrophy of turbot Scophthalmus maximus by dietary Clostridium autoethanogenum protein

Aquaculture Reports, Journal Year: 2025, Volume and Issue: 40, P. 102603 - 102603

Published: Jan. 2, 2025

Language: Английский

Evidence for Interaction of 5,10-Methylenetetrahydrofolate Reductase (MTHFR) with Methylenetetrahydrofolate Dehydrogenase (MTHFD1) and General Control Nonderepressible 1 (GCN1)

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 22, 2024

5,10-methylenetetrahydrofolate reductase (MTHFR) is a folate cycle enzyme required for the intracellular synthesis of methionine. MTHFR was previously shown to be partially phosphorylated at 16 residues, which abrogated by conversion threonine 34 alanine (T34A) or truncation first 37 amino acids (i.e. expression 38-656), and promoted methionine supplementation. Here, we over-expressed wild-type (MTFHRWT), as well variants MTHFRT34A MTHFR38-656 in 293T cells provide further insights into these mechanisms. We demonstrate that following incubation high conditions (100-1000 uM) MTHFRWT almost completely phosphorylated, but restricted (0-10 phosphorylation reduced, while always remains unphosphorylated. Following affinity purification coupled mass spectrometry an empty vector, MTHFRWT, three separate experiments, identified 134 proteins consistently pulled-down all protein variants, 5 were indicated likely true interactors (SAINT prediction threshold 0.95 2 fold-change). Amongst methylenetetrahydrofolate dehydrogenase (MTHFD1) acid starvation sensor General Control Nonderepressible 1 (GCN1). Immunoprecipitation-immunoblotting replicated interaction with both proteins. An AlphaFold 3 generated model MTHFR-MTHFD1 places MTHFD1 dehydrogenase/cyclohydrolase domain direct contact catalytic domain, suggesting their may facilitate delivery methylenetetrahydrofolate. Overall, confirm availability increases phosphorylation, potential GCN1.

Language: Английский

Evidence for Interaction of 5,10-Methylenetetrahydrofolate Reductase (MTHFR) with Methylenetetrahydrofolate Dehydrogenase (MTHFD1) and General Control Nonderepressible 1 (GCN1)

Biochimie, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 1, 2024

5,10-methylenetetrahydrofolate reductase (MTHFR) is a folate cycle enzyme required for the intracellular synthesis of methionine. MTHFR was previously shown to be partially phosphorylated at 16 residues, which abrogated by conversion threonine 34 alanine (T34A) or truncation first 37 amino acids (i.e. expression 38-656), and promoted methionine supplementation. Here, we over-expressed wild-type (MTFHR

Language: Английский