Purification, Characterization, and Fractionation of the δ-Aminolevulinic Acid Synthesizing Enzymes from Light-Grown Chlamydomonas reinhardtii Cells
Open Access
- 1 March 1984
- journal article
- research article
- Published by Oxford University Press (OUP) in Plant Physiology
- Vol. 74 (3) , 569-575
- https://doi.org/10.1104/pp.74.3.569
Abstract
The synthesis of δ-aminolevulinate from glutamate by Chlamydomonas reinhardtii membrane-free cell homogenates requires Mg2+, ATP, and NADPH as cofactors. The pH optimum is about 8.3. When analyzed by a Fractogel TSK gel filtration column the δ-aminolevulinate synthesizing enzymes, including glutamate-1-semialdehyde aminotransferase, elute with an apparent molecular weight of about 45,000. The enzymes obtained from the gel filtration column were separated into three fractions by affinity column chromatography. One fraction binds to heme-Sepharose, one to Blue Sepharose, while the enzyme converting the putative glutamate-1-semialdehyde to δ-aminolevulinic acid is retained by neither column. All three fractions are necessary for the conversion of glutamate to δ-aminolevulinate. The δ-aminolevulinate synthesizing enzymes from Chlamydomonas are sensitive to inhibition by heme but not sensitive to inhibition by protoporphyrin.Keywords
This publication has 22 references indexed in Scilit:
- The Proof by 13C‐NMR Spectroscopy of the Predominance of the C5 Pathway over the Shemin Pathway in Chlorophyll Biosynthesis in Higher Plants and of the Formation of the Methyl Ester Group of Chlorophyll from GlycineEuropean Journal of Biochemistry, 1983
- Formation of δ-aminolevulinic acid from glutamic acid by a partially purified enzyme system from wheat leavesBiochimica et Biophysica Acta (BBA) - Enzymology, 1979
- Mg-protoporphyrin-IX and δ-aminolevulinic acid synthesis from glutamate in isolated greening chloroplastsArchives of Biochemistry and Biophysics, 1978
- A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye BindingAnalytical Biochemistry, 1976
- Biosynthesis of δ-aminolevulinic acid in the unicellular rhodophyte, cyanidium caldariumBiochemical and Biophysical Research Communications, 1976
- Biosynthesis of delta-aminolevulinic acid from the intact carbon skeleton of glutamic acid in greening barley.Proceedings of the National Academy of Sciences, 1975
- GENETIC CONTROL OF CHLOROPHYLL BIOSYNTHESIS IN CHLAMYDOMONAS The Journal of cell biology, 1974
- Defective synthesis of porphyrins in barley plastids caused by mutation in nuclear genesBiochimica et Biophysica Acta (BBA) - General Subjects, 1972
- Assays for Porphyrins, δ-Aminolevulinic-Acid Dehydratase, and Porphyrinogen Synthetase in Microliter Samples of Whole Blood: Applications to Metabolic Defects Involving the Heme PathwayProceedings of the National Academy of Sciences, 1972
- Dithiothreitol, a New Protective Reagent for SH Groups*Biochemistry, 1964