RGD Reference Report - Assays of methylenetetrahydrofolate reductase and methionine synthase activities by monitoring 5-methyltetrahydrofolate and tetrahydrofolate using high-performance liquid chromatography with fluorescence detection. - Rat Genome Database

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Assays of methylenetetrahydrofolate reductase and methionine synthase activities by monitoring 5-methyltetrahydrofolate and tetrahydrofolate using high-performance liquid chromatography with fluorescence detection.

Authors: Huang, L  Zhang, J  Hayakawa, T  Tsuge, H 
Citation: Huang L, etal., Anal Biochem. 2001 Dec 15;299(2):253-9.
RGD ID: 1601423
Pubmed: PMID:11730351   (View Abstract at PubMed)
DOI: DOI:10.1006/abio.2001.5421   (Journal Full-text)

We developed a method for assays of methylenetetrahydrofolate reductase and methionine synthase activities by monitoring their products of 5-methyltetrahydrofolate (5-CH(3)-H(4)folate) and tetrahydrofolate (H(4)folate) directly, using high-performance liquid chromatography with fluorescence detection. Folate derivatives and enzymes were stable in the assay process. No reagents in the assay mixture were found to disturb the separation and detection of both H(4)folate and 5-CH(3)-H(4)folate in our assay system. The detection limit of this method was less than 20 nM H(4)folate or 5-CH(3)-H(4)folate in the enzyme assay system. This analytical method, therefore, has a sensitivity high enough to obtain accurate parameters of Michaelis-Menten kinetics and for assays of crude extracts from various biological samples. In addition, the analytical procedure is very simple and economical; it may be a useful tool for studying methylenetetrahydrofolate reductase and methionine synthase activities.



Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
MtrRathomocysteine metabolic process  IDA  RGD 
MtrRatmethionine biosynthetic process  IDA  RGD 
MthfrRattetrahydrofolate metabolic process  IDA  RGD 
MtrRattetrahydrofolate metabolic process  IDA  RGD 

Molecular Function

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
MtrRatamino acid binding  IDA  RGD 
MthfrRatflavin adenine dinucleotide binding  IDA  RGD 
MtrRatfolic acid binding  IDA  RGD 
MtrRatmethionine synthase activity  IDA  RGD 
MthfrRatmethylenetetrahydrofolate reductase (NAD(P)H) activity  IDA  RGD 
MthfrRatNADP binding  IDA  RGD 

Objects Annotated

Genes (Rattus norvegicus)
Mthfr  (methylenetetrahydrofolate reductase)
Mtr  (5-methyltetrahydrofolate-homocysteine methyltransferase)


Additional Information