Enzymatic Assay for Adenosylcobalamin-dependent Methylmalonyl Coenzyme A Mutase
نویسندگان
چکیده
منابع مشابه
Protein-coenzyme interactions in adenosylcobalamin-dependent glutamate mutase.
Glutamate mutase catalyses an unusual isomerization involving free-radical intermediates that are generated by homolysis of the cobalt-carbon bond of the coenzyme adenosylcobalamin (coenzyme B(12)). A variety of techniques have been used to examine the interaction between the protein and adenosylcobalamin, and between the protein and the products of coenzyme homolysis, cob(II)alamin and 5'-deox...
متن کاملFully automated assay for cobalamin-dependent methylmalonyl CoA mutase.
We constructed a fully automated assay for the cobalamin-dependent enzyme methylmalonyl coenzyme A (CoA) mutase. The assay involves preincubation of the enzyme with adenosylcobalamin, incubation with substrate, termination of the reaction by adding trichloroacetic acid, filtration to remove precipitated protein, and finally analysis of the filtrate (containing methylmalonyl CoA and the product ...
متن کاملThe Absolute Configuration of Methylmalonyl Coenzyme A and Stereochemistry of the Methylmalonyl Coenzyme A Mutase Reaction*
Rropionyl coenzyme A was carboxylated with epimerasefree carboxylase, and the resulting methylmalonyl coenzyme A (a) was reduced with Raney nickel. Isolation of (-)(R)-P-hydroxyisobutyric acid N-phenylcarbamate (11) from the reaction mixture showed that methylmalonyl coenzyme A (a) has the (S) configuration. Propionyl coenzyme A was also converted to deuteriosuccinate with a DrO extract of a be...
متن کاملAdenosylcobalamin-dependent methylmalonyl-CoA mutase from Propionibacterium shermanii. Active holoenzyme produced from Escherichia coli.
The linked structural genes coding for both subunits of adenosylcobalamin-dependent methylmalonyl-CoA mutase from the Gram-positive bacterium Propionibacterium shermanii have been altered by site-directed mutagenesis and placed under the control of an inducible phage-T7-specific plasmid promoter in Escherichia coli. Conditions have been found under which both alpha- and beta-subunits are produc...
متن کاملActivation of methylmalonyl-Coenzyme A mutase by MeaB
In mammalian cells, only two enzymes are known to require vitamin B12 (cobalamin, Cbl) derivatives as a cofactor: methionine synthase, a cytosolic enzyme, which uses methylcobalamin, and methylmalonyl-coenzyme A mutase (MCM), a mitochondrial enzyme, which uses 5’-deoxyadenosylcobalamin (AdoCbl) (1). The latter enzyme has a broad distribution among living organisms and is found in both bacteria ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Bioscience, Biotechnology, and Biochemistry
سال: 1993
ISSN: 0916-8451,1347-6947
DOI: 10.1271/bbb.57.1593