Skip to main content
PMC home page
Plant Physiology logoLink to Plant Physiology
. 1993 Nov;103(3):943–948. doi: 10.1104/pp.103.3.943

Identification of Distinct Internal and External Isozymes of Carbonic Anhydrase in Chlorella saccharophila.

T G Williams 1, B Colman 1
PMCID: PMC159067  PMID: 12231991

Abstract

External carbonic anhydrase (CA) was detected in whole cells of alkaline-grown Chlorella saccharophila but was suppressed by growth at acid pH or growth on elevated levels of CO2. Internal CA activity was measured potentiometrically as an increase in activity in cell extracts over that of intact cells. Cells grown under all conditions had equal levels of internal CA activity. Two isozymes were identified after electrophoretic separation of soluble proteins on cellulose acetate plates. The fast isozyme was found in cells grown under all conditions, whereas the slow isozyme was found only in cells grown at alkaline pH. Western blot analysis following sodium dodecyl sulfate-polyacrylamide gel electrophoresis using antibodies produced against the periplasmic form of CA from Chlamydomonas reinhardtii revealed a single band at 39 kD, which did not change in intensity between growth conditions and was associated only with proteins eluted from the fast band. The slow isozyme was inactivated by incubation of cell extract at 30[deg]C and by incubation in 10 mM dithiothreitol, whereas the internal form was unaffected. These results indicate that external and internal forms of CA differ in structure and their activities respond differently to environmental conditions.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Burnell J. N., Hatch M. D. Low bundle sheath carbonic anhydrase is apparently essential for effective c(4) pathway operation. Plant Physiol. 1988 Apr;86(4):1252–1256. doi: 10.1104/pp.86.4.1252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Coleman J. R., Rotatore C., Williams T. G., Colman B. Identification and localization of carbonic anhydrase in two chlorella species. Plant Physiol. 1991 Jan;95(1):331–334. doi: 10.1104/pp.95.1.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fawcett T. W., Browse J. A., Volokita M., Bartlett S. G. Spinach carbonic anhydrase primary structure deduced from the sequence of a cDNA clone. J Biol Chem. 1990 Apr 5;265(10):5414–5417. [PubMed] [Google Scholar]
  4. Fukuzawa H., Suzuki E., Komukai Y., Miyachi S. A gene homologous to chloroplast carbonic anhydrase (icfA) is essential to photosynthetic carbon dioxide fixation by Synechococcus PCC7942. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4437–4441. doi: 10.1073/pnas.89.10.4437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Graham D., Reed M. L., Patterson B. D., Hockley D. G., Dwyer M. R. Chemical properties, distribution, and physiology of plant and algal carbonic anhydrases. Ann N Y Acad Sci. 1984;429:222–237. doi: 10.1111/j.1749-6632.1984.tb12340.x. [DOI] [PubMed] [Google Scholar]
  6. Guilloton M. B., Korte J. J., Lamblin A. F., Fuchs J. A., Anderson P. M. Carbonic anhydrase in Escherichia coli. A product of the cyn operon. J Biol Chem. 1992 Feb 25;267(6):3731–3734. [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Majeau N., Coleman J. R. Isolation and characterization of a cDNA coding for pea chloroplastic carbonic anhydrase. Plant Physiol. 1991 Jan;95(1):264–268. doi: 10.1104/pp.95.1.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Moroney J. V., Togasaki R. K., Husic H. D., Tolbert N. E. Evidence That an Internal Carbonic Anhydrase Is Present in 5% CO(2)-Grown and Air-Grown Chlamydomonas. Plant Physiol. 1987 Jul;84(3):757–761. doi: 10.1104/pp.84.3.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Price G. D., Coleman J. R., Badger M. R. Association of Carbonic Anhydrase Activity with Carboxysomes Isolated from the Cyanobacterium Synechococcus PCC7942. Plant Physiol. 1992 Oct;100(2):784–793. doi: 10.1104/pp.100.2.784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Williams T. G., Turpin D. H. The Role of External Carbonic Anhydrase in Inorganic Carbon Acquisition by Chlamydomonas reinhardii at Alkaline pH. Plant Physiol. 1987 Jan;83(1):92–96. doi: 10.1104/pp.83.1.92. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES