In vitro synthesis of the pyruvate dehydrogenase complex components of Ascaris suum mitochondria

Conference · Fri May 01 00:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6023723

Desai, S; Ruff, V; DuBrul, E F; Komuniecki, R W

The pyruvate dehydrogenase complex (PDC) plays a pivotal role in the anaerobic metabolism of Ascaris suum mitochondria. They have initiated a series of studies on the in vitro synthesis and mitochondrial import of PDC. PDC has been purified from adult Ascaris body wall muscle, fully phosphorylated in vitro, and separated into its component subunits on SDS/PAGE. The individual components were electroeluted from the gels and used to immunize rabbits. IgG's to the individual subunits were prepared from antisera and their specificities were verified by immuno-blotting. Each IgG identified a single specific band at the appropriate location in extracts of adult Ascaris body wall muscle mitochondria. Poly A/sup +/-RNA was prepared from body wall muscle and translated in a reticylocyte lysate system using /sup 35/S-methionine. Translation products were immunoprecipitated with specific IgG's, electrophoresed, and fluorographed. Each immunoprecipitation gave rise to a single radioactive polypeptide that was slightly larger than the specific PDC subunit isolated from the adult mitochondria. This system has demonstrated its feasibility for the study of mitochondrial import of a multienzyme complex that is critical for the anaerobic mitochondrial metabolism of Ascaris suum.

• Research Organization: Univ. of Toledo, OH
• OSTI ID: 6023723
• Report Number(s): CONF-870644-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 46:6; ISSN FEPRA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ASCARIDAE
ASCARIS
ASCHELMINTHES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTROPHORESIS
ENZYMES
EVEN-ODD NUCLEI
FRACTIONATION
HELMINTHS
IN VITRO
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
METABOLISM
METHIONINE
MITOCHONDRIA
NEMATODES
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANOIDS
OXIDOREDUCTASES
PARASITES
PHOSPHORYLATION
RADIOISOTOPES
SEPARATION PROCESSES
SULFUR 35
SULFUR ISOTOPES
TRACER TECHNIQUES