delta-Aminolevulinic acid synthetase from rat liver mitochondria. Purification and properties

delta-Aminolevulinic acid synthetase has been purified from liver mitochondria of young, uninduced rats. After nonionic detergent solubilization of mitochondrial inner membrane-matrix fractions, the enzyme was purified to a specific activity of approximately 2,000 nmol of delta-aminolevulinic acid f...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1979-07, Vol.254 (13), p.6112-6118
Main Authors: J R Paterniti, Jr, D S Beattie
Format: Article
Language:English
Subjects:
5-Aminolevulinate Synthetase - isolation & purification
5-Aminolevulinate Synthetase - metabolism
Animals
Hemin - pharmacology
Kinetics
Macromolecular Substances
Male
Mitochondria, Liver - enzymology
Molecular Weight
Rats
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:delta-Aminolevulinic acid synthetase has been purified from liver mitochondria of young, uninduced rats. After nonionic detergent solubilization of mitochondrial inner membrane-matrix fractions, the enzyme was purified to a specific activity of approximately 2,000 nmol of delta-aminolevulinic acid formed/h/mg of protein at 30 degrees C, by means of ammonium sulfate precipitation, diethylaminoethyl cellulose chromatography, Sephacryl chromatography, and preparative gel electrophoresis. The purified enzyme preparation thus obtained was apparently homogeneous as judged by its migration as a single band with a molecular weight of 58,000 +/- 6,000 upon electrophoresis in sodium dodecyl sulfate polyacrylamide gels. The native enzyme probably exists as a dimer with a molecular weight of approximately 120,000. A pH optimum of 7.5 and an isoelectric point of 4.5 were also determined. Both monovalent cations and hemin strongly inhibited the activity of the purified enzyme.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)50526-3