Properties of chicken erythrocyte delta-aminolevulinate synthase.

Abstract

A procedure is described for preparing a fraction highly enriched for chicken blood delta-aminolevulinate synthase (ALA-S) using animals recovering from acetylphenylhydrazine-induced anemia. 1. Blood cells collected from chickens recovering from anemia were disrupted by nitrogen cavitation, and the mitochondrial fraction was prepared from the cell homogenates. ALA-S was released then from mitochondria by sonication and isolated by a procedure involving gel filtration chromatography on Sephadex G-150, fractionation with ammonium sulfate, ion exchange chromatography on DEAE-Sephacel, and preparative isoelectric focusing. 2. Electrophoretic analyses under denaturing conditions indicated that the final ALA-S preparation was particularly enriched from a 62,200 Da polypeptide. The enzyme eluted from Sephadex G-200 with an equivalent molecular weight of 115,000; this suggested that active ALA-S was a dimer. 3. ALA-S was most active in the pH range of 7.0-8.0, with an apparent KM of 13 microM for succinyl-CoA and of 4.0 mM for glycine. The activity was inhibited 50% by 30 microM hemin.

• Authors: I Z Ades, D M Friedland
• Journal: The International journal of biochemistry (Int J Biochem) Vol. 20 Issue 9 Pg. 965-9 ( 1988) ISSN: 0020-711X [Print] England
• PMID: 3197910 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Phenylhydrazines
• N(1)-acetylphenylhydrazine
• 5-Aminolevulinate Synthetase

Topics

• 5-Aminolevulinate Synthetase (blood)
• Anemia (chemically induced)
• Animals
• Cell Fractionation
• Chickens
• Erythrocytes (enzymology)
• Hydrogen-Ion Concentration
• Isoelectric Focusing
• Molecular Weight
• Phenylhydrazines