Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1994 May;105(1):133–139. doi: 10.1104/pp.105.1.133

Identification of Intracellular Carbonic Anhydrase in Chlamydomonas reinhardtii with a Carbonic Anhydrase-Directed Photoaffinity Label.

H D Husic 1, C A Marcus 1
PMCID: PMC159338  PMID: 12232193

Abstract

A carbonic anhydrase (CA)-directed photoaffinity reagent, 125I-labeled p-aminomethylbenzenesulfonamide-4-azidosalicylamide,was synthesized and shown to derivatize periplasmic CA in the unicellular green alga Chlamydomonas reinhardtii. The photoderivatization of purified C. reinhardtii periplasmic CA or intact C. reinhardtii cells with the reagent resulted in the modification of the large (37 kD) subunit of the enzyme. Photoderivatization of proteins in lysed C. reinhardtii cells also resulted in the specific labeling of a polypeptide of 30 kD. Centrifugation of the cell extract prior to photoaffinity labeling revealed that the labeled peptide was present predominantly in a particulate fraction. The photoaffinity-labeled 30-kD polypeptide was not observed in extracts from a mutant of C. reinhardtii that is believed to be deficient in an intracellular form of CA. These results provide evidence that the 30-kD polypeptide, which is photoaffinity labeled in lysed C. reinhardtii cells, is an intracellular form of CA.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bayley H., Knowles J. R. Photoaffinity labeling. Methods Enzymol. 1977;46:69–114. doi: 10.1016/s0076-6879(77)46012-9. [DOI] [PubMed] [Google Scholar]
  2. Coleman J. R., Berry J. A., Togasaki R. K., Grossman A. R. Identification of Extracellular Carbonic Anhydrase of Chlamydomonas reinhardtii. Plant Physiol. 1984 Oct;76(2):472–477. doi: 10.1104/pp.76.2.472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Husic H. D., Hsieh S., Berrier A. L. Effect of dithiothreitol on the catalytic activity, quaternary structure and sulfonamide-binding properties of an extracellular carbonic anhydrase from Chlamydomonas reinhardtii. Biochim Biophys Acta. 1991 May 30;1078(1):35–42. doi: 10.1016/0167-4838(91)90089-i. [DOI] [PubMed] [Google Scholar]
  4. Husic H. D., Kitayama M., Togasaki R. K., Moroney J. V., Morris K. L., Tolbert N. E. Identification of Intracellular Carbonic Anhydrase in Chlamydomonas reinhardtii which Is Distinct from the Periplasmic Form of the Enzyme. Plant Physiol. 1989 Mar;89(3):904–909. doi: 10.1104/pp.89.3.904. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kamo T., Shimogawara K., Fukuzawa H., Muto S., Miyachi S. Subunit constitution of carbonic anhydrase from Chlamydomonas reinhardtii. Eur J Biochem. 1990 Sep 11;192(2):557–562. doi: 10.1111/j.1432-1033.1990.tb19261.x. [DOI] [PubMed] [Google Scholar]
  6. Kandel S. I., Wong S. C., Kandel M., Gornall A. G. Irreversible inactivation of bovine carbonic anhydrase B by bromoacetazolamide. J Biol Chem. 1968 May 10;243(9):2437–2439. [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Maren T. H. Carbonic anhydrase: chemistry, physiology, and inhibition. Physiol Rev. 1967 Oct;47(4):595–781. doi: 10.1152/physrev.1967.47.4.595. [DOI] [PubMed] [Google Scholar]
  9. Moroney J. V., Husic H. D., Tolbert N. E. Effect of Carbonic Anhydrase Inhibitors on Inorganic Carbon Accumulation by Chlamydomonas reinhardtii. Plant Physiol. 1985 Sep;79(1):177–183. doi: 10.1104/pp.79.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Rawat M., Moroney J. V. Partial characterization of a new isoenzyme of carbonic anhydrase isolated from Chlamydomonas reinhardtii. J Biol Chem. 1991 May 25;266(15):9719–9723. [PubMed] [Google Scholar]
  11. Spalding M. H., Spreitzer R. J., Ogren W. L. Carbonic Anhydrase-Deficient Mutant of Chlamydomonas reinhardii Requires Elevated Carbon Dioxide Concentration for Photoautotrophic Growth. Plant Physiol. 1983 Oct;73(2):268–272. doi: 10.1104/pp.73.2.268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sueoka N. MITOTIC REPLICATION OF DEOXYRIBONUCLEIC ACID IN CHLAMYDOMONAS REINHARDI. Proc Natl Acad Sci U S A. 1960 Jan;46(1):83–91. doi: 10.1073/pnas.46.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sültemeyer D. F., Fock H. P., Canvin D. T. Mass Spectrometric Measurement of Intracellular Carbonic Anhydrase Activity in High and Low C(i) Cells of Chlamydomonas: Studies Using O Exchange with C/O Labeled Bicarbonate. Plant Physiol. 1990 Nov;94(3):1250–1257. doi: 10.1104/pp.94.3.1250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. van der Horst G. T., Mancini G. M., Brossmer R., Rose U., Verheijen F. W. Photoaffinity labeling of a bacterial sialidase with an aryl azide derivative of sialic acid. J Biol Chem. 1990 Jul 5;265(19):10801–10804. [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES