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Title: Evidence for chloroplastic succinate dehydrogenase participating in the chloroplastic respiratory and photosynthetic electron transport chains of Chlamydomonas reinhardtii

Abstract

A method for isolating intact chloroplasts from Chlamydomonas reinhardtii F-60 was developed from the Klein, Chen, Gibbs, Platt-Aloia procedure. Protoplasts, generated by treatment with autolysine, were lysed with a solution of digitonin and fractionated on Percoll step gradients. The chloroplasts were assessed to be 90% intact (ferricyanide assay) and free from cytoplasmic contamination (NADP isocitrate dehydrogenase activity) and to range from 2 to 5% in mitochondrial contamination (cytochrome c oxidase activity). About 25% of the cellular succinate dehydrogenase activity (21.6 micromoles per milligram chlorophyll per hour, as determined enzymically) was placed within the chloroplast. Chloroplastic succinate dehydrogenase had a K{sub m} for succinate of 0.55 millimolar and was associated with the thylakoidal material derived from the intact chloroplasts. This same thylakoidal material, with an enzymic assay of 21.6 micromoles per milligram chlorophyll per hour was able to initiate a light-dependent uptake of oxygen at a rate of 16.4 micromoles per milligram chlorophyll per hour when supplied with succinate and methyl viologen. Malonate was an apparent competitive inhibitor of this reaction. The succinate dehydrogenase activity present in the chloroplast was sufficient to account for the photoanaerobic rate of acetate dissimilation in H{sub 2} adapted Chlamydomonas.

Authors:
; ;  [1]
  1. Brandeis Univ., Waltham, MA (USA)
Publication Date:
OSTI Identifier:
7151146
DOE Contract Number:  
AC02-76ER03231
Resource Type:
Journal Article
Journal Name:
Plant Physiology; (USA)
Additional Journal Information:
Journal Volume: 90:3; Journal ID: ISSN 0032-0889
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CHLOROPLASTS; ENZYME ACTIVITY; ACETATES; CHLAMYDOMONAS; OXIDATION; OXIDOREDUCTASES; PHOTOSYNTHESIS; SUCCINIC ACID; ALGAE; CARBOXYLIC ACID SALTS; CARBOXYLIC ACIDS; CELL CONSTITUENTS; CHEMICAL REACTIONS; CHLOROPHYCOTA; DICARBOXYLIC ACIDS; ENZYMES; MICROORGANISMS; ORGANIC ACIDS; ORGANIC COMPOUNDS; PHOTOCHEMICAL REACTIONS; PLANTS; SYNTHESIS; UNICELLULAR ALGAE; 551000* - Physiological Systems

Citation Formats

Willeford, K O, Gombos, Z, and Gibbs, M. Evidence for chloroplastic succinate dehydrogenase participating in the chloroplastic respiratory and photosynthetic electron transport chains of Chlamydomonas reinhardtii. United States: N. p., 1989. Web. doi:10.1104/pp.90.3.1084.
Willeford, K O, Gombos, Z, & Gibbs, M. Evidence for chloroplastic succinate dehydrogenase participating in the chloroplastic respiratory and photosynthetic electron transport chains of Chlamydomonas reinhardtii. United States. https://doi.org/10.1104/pp.90.3.1084
Willeford, K O, Gombos, Z, and Gibbs, M. 1989. "Evidence for chloroplastic succinate dehydrogenase participating in the chloroplastic respiratory and photosynthetic electron transport chains of Chlamydomonas reinhardtii". United States. https://doi.org/10.1104/pp.90.3.1084.
@article{osti_7151146,
title = {Evidence for chloroplastic succinate dehydrogenase participating in the chloroplastic respiratory and photosynthetic electron transport chains of Chlamydomonas reinhardtii},
author = {Willeford, K O and Gombos, Z and Gibbs, M},
abstractNote = {A method for isolating intact chloroplasts from Chlamydomonas reinhardtii F-60 was developed from the Klein, Chen, Gibbs, Platt-Aloia procedure. Protoplasts, generated by treatment with autolysine, were lysed with a solution of digitonin and fractionated on Percoll step gradients. The chloroplasts were assessed to be 90% intact (ferricyanide assay) and free from cytoplasmic contamination (NADP isocitrate dehydrogenase activity) and to range from 2 to 5% in mitochondrial contamination (cytochrome c oxidase activity). About 25% of the cellular succinate dehydrogenase activity (21.6 micromoles per milligram chlorophyll per hour, as determined enzymically) was placed within the chloroplast. Chloroplastic succinate dehydrogenase had a K{sub m} for succinate of 0.55 millimolar and was associated with the thylakoidal material derived from the intact chloroplasts. This same thylakoidal material, with an enzymic assay of 21.6 micromoles per milligram chlorophyll per hour was able to initiate a light-dependent uptake of oxygen at a rate of 16.4 micromoles per milligram chlorophyll per hour when supplied with succinate and methyl viologen. Malonate was an apparent competitive inhibitor of this reaction. The succinate dehydrogenase activity present in the chloroplast was sufficient to account for the photoanaerobic rate of acetate dissimilation in H{sub 2} adapted Chlamydomonas.},
doi = {10.1104/pp.90.3.1084},
url = {https://www.osti.gov/biblio/7151146}, journal = {Plant Physiology; (USA)},
issn = {0032-0889},
number = ,
volume = 90:3,
place = {United States},
year = {Sat Jul 01 00:00:00 EDT 1989},
month = {Sat Jul 01 00:00:00 EDT 1989}
}