Molecular cloning of L-methylmalonyl-CoA mutase: Gene transfer and analysis of mut cell lines

Journal Article · Sun May 01 00:00:00 EDT 1988 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.85.10.3518· OSTI ID:5406224

Ledley, F D; Lumetta, M; Nguyen, P N; Kolhouse, J F; Allen, R H ^[1]

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1. Baylor College of Medicine, Houston, TX (USA)

L-Methylmalonyl-CoA mutase (MCM, EC 5.4.99.2) is a mitochondrial adenosylcobalamin-requiring enzyme that catalyzes the isomerization of L-methylmalonyl-CoA to succinyl-CoA. This enzyme is deficient in methylmalonic acidemia, an often fatal disorder of organic acid metabolism. Antibody against human placental MCM was used to screen human placenta and liver cDNA expression libraries for MCM cDNA clones. One clone expressed epitopes that could affinity-purify antibodies against MCM. A cDNA corresponding in length to the mRNA was obtained and introduced into COS cells by DNA-mediated gene transfer. Cells transformed with this clone expressed increased levels of MCM enzymatic activity. RNA blot analysis of cells genetically deficient in MCM indicates that several deficient cell lines have a specific decrease in the amount of hybridizable mRNA. These data confirm the authenticity of the MCM cDNA clone, establish the feasibility of constituting MCM activity by gene transfer for biochemical analysis and gene therapy, and provide a preliminary picture of the genotypic spectrum underlying MCM deficiency.

• OSTI ID: 5406224
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 85:10; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ENZYME ACTIVITY
HEREDITARY DISEASES
ETIOLOGY
ISOMERASES
DNA-CLONING
ANTIBODIES
IODINE 125
LIVER
MAN
MESSENGER-RNA
METABOLISM
PLACENTA
RECOMBINANT DNA
ANIMALS
BETA DECAY RADIOISOTOPES
BODY
CLONING
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
DISEASES
DNA
DNA HYBRIDIZATION
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
FETAL MEMBRANES
GLANDS
HYBRIDIZATION
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPES
MAMMALS
MEMBRANES
NUCLEI
NUCLEIC ACIDS
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PRIMATES
RADIOISOTOPES
RNA
VERTEBRATES
550201* - Biochemistry- Tracer Techniques