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. 1986 Feb;38(2):125–136.

Carbonic anhydrase II deficiency: diagnosis and carrier detection using differential enzyme inhibition and inactivation.

V Sundaram, P Rumbolo, J Grubb, P Strisciuglio, W S Sly
PMCID: PMC1684750  PMID: 3080873

Abstract

Carbonic anhydrase (CA) I and II are soluble isozymes that represent the major nonhemoglobin proteins in the erythrocyte. We recently identified a deficiency of CA II as the enzymatic basis for the autosomal recessive syndrome of osteopetrosis with renal tubular acidosis and cerebral calcification. Virtual absence of the CA II peak on high-performance liquid chromatography, of CA II esterase activity, and of immunoprecipitable CA II were demonstrated on extracts of red cell lysates from all patients studied. Reduced levels of CA II were found in obligate heterozygotes. Here, we present evidence that CA II in red cell lysates can be quantitated by measuring CO2 hydratase activity in the presence of inhibitors that selectively inhibit the activity of CA I to a much greater extent than that of CA II. This was done with iodide (anion binding) and bromopyruvic acid (alkylation), and the respective assays evaluated as diagnostic tools for CA II deficiency in human red cells. These techniques greatly simplify the quantitation of CA II in hemolysates and should make genetic diagnosis and counseling for the newly described inborn error of metabolism due to CA II deficiency generally available. They also allow quantitation of CA I in red cell lysates.

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Selected References

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

  1. Carter N. D., Heath R., Welty R. J., Hewett-Emmett D., Jeffery S., Shiels A., Tashian R. E. Red cells genetically deficient in carbonic anhydrase II have elevated levels of a carbonic anhydrase indistinguishable from muscle CA III. Ann N Y Acad Sci. 1984;429:284–286. doi: 10.1111/j.1749-6632.1984.tb12351.x. [DOI] [PubMed] [Google Scholar]
  2. DeSimone J., Magid E., Tashian R. E. Genetic variation in the carbonic anhydrase isozymes of macaque monkeys. II. Inheritance of red cell carbonic anhydrase levels in different carbonic anhydrase I genotypes of the pig-tailed macaque, Macaca nemestrina. Biochem Genet. 1973 Feb;8(2):165–174. doi: 10.1007/BF00485544. [DOI] [PubMed] [Google Scholar]
  3. Ferrell R. E., Osborne W. R., Tashian R. E. Effect of metabolic acidosis on hydrogen ion excretion in a pigtail macaque with erythrocyte carbonic anhydrase I deficiency. Proc Soc Exp Biol Med. 1981 Nov;168(2):155–158. doi: 10.3181/00379727-168-41251. [DOI] [PubMed] [Google Scholar]
  4. Guibaud P., Larbre F., Freycon M. T., Genoud J. Ostéopétrose et acidose rénale tubulaire. Deux cas de cette association dans une fratrie. Arch Fr Pediatr. 1972 Mar;29(3):269–286. [PubMed] [Google Scholar]
  5. Göthe P. O., Nyman P. O. Inactivation of human erythrocyte carbonic anhydrases by bromopyruvate. FEBS Lett. 1972 Mar 15;21(2):159–164. doi: 10.1016/0014-5793(72)80127-3. [DOI] [PubMed] [Google Scholar]
  6. Heath R., Jeffery S., Carter N. Radioimmunoassay of human muscle carbonic anhydrase III in dystrophic states. Clin Chim Acta. 1982 Mar 12;119(3):299–305. doi: 10.1016/0009-8981(82)90343-6. [DOI] [PubMed] [Google Scholar]
  7. Kendall A. G., Tashian R. E. Erythrocyte carbonic anhydrase I: inherited deficiency in humans. Science. 1977 Jul 29;197(4302):471–472. doi: 10.1126/science.406674. [DOI] [PubMed] [Google Scholar]
  8. MAREN T. H. A simplified micromethod for the determination of carbonic anhydrase and its inhibitors. J Pharmacol Exp Ther. 1960 Sep;130:26–29. [PubMed] [Google Scholar]
  9. 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]
  10. Maren T. H., Couto E. O. The nature of anion inhibition of human red cell carbonic anhydrases. Arch Biochem Biophys. 1979 Sep;196(2):501–510. doi: 10.1016/0003-9861(79)90302-3. [DOI] [PubMed] [Google Scholar]
  11. McKinley D. N., Whitney P. L. Particulate carbonic anhydrase in homogenates of human kidney. Biochim Biophys Acta. 1976 Oct 11;445(3):780–790. doi: 10.1016/0005-2744(76)90128-5. [DOI] [PubMed] [Google Scholar]
  12. Sanyal G., Pessah N. I., Maren T. H. Kinetics and inhibition of membrane-bound carbonic anhydrase from canine renal cortex. Biochim Biophys Acta. 1981 Jan 15;657(1):128–137. doi: 10.1016/0005-2744(81)90136-4. [DOI] [PubMed] [Google Scholar]
  13. Sly W. S., Hewett-Emmett D., Whyte M. P., Yu Y. S., Tashian R. E. Carbonic anhydrase II deficiency identified as the primary defect in the autosomal recessive syndrome of osteopetrosis with renal tubular acidosis and cerebral calcification. Proc Natl Acad Sci U S A. 1983 May;80(9):2752–2756. doi: 10.1073/pnas.80.9.2752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sly W. S., Whyte M. P., Sundaram V., Tashian R. E., Hewett-Emmett D., Guibaud P., Vainsel M., Baluarte H. J., Gruskin A., Al-Mosawi M. Carbonic anhydrase II deficiency in 12 families with the autosomal recessive syndrome of osteopetrosis with renal tubular acidosis and cerebral calcification. N Engl J Med. 1985 Jul 18;313(3):139–145. doi: 10.1056/NEJM198507183130302. [DOI] [PubMed] [Google Scholar]
  15. Tashian R. E., Hewett-Emmett D., Dodgson S. J., Forster R. E., 2nd, Sly W. S. The value of inherited deficiencies of human carbonic anhydrase isozymes in understanding their cellular roles. Ann N Y Acad Sci. 1984;429:262–275. doi: 10.1111/j.1749-6632.1984.tb12346.x. [DOI] [PubMed] [Google Scholar]
  16. Tashian R. E., Hewett-Emmett D., Goodman M. On the evolution and genetics of carbonic anhydrases I, II, and III. Isozymes Curr Top Biol Med Res. 1983;7:79–100. [PubMed] [Google Scholar]
  17. Vainsel M., Fondu P., Cadranel S., Rocmans C., Gepts W. Osteopetrosis associated with proximal and distal tubular acidosis. Acta Paediatr Scand. 1972 Jul;61(4):429–434. doi: 10.1111/j.1651-2227.1972.tb15859.x. [DOI] [PubMed] [Google Scholar]
  18. Venta P. J., Montgomery J. C., Wiebauer K., Hewett-Emmett D., Tashian R. E. Organization of the mouse and human carbonic anhydrase II genes. Ann N Y Acad Sci. 1984;429:309–323. doi: 10.1111/j.1749-6632.1984.tb12355.x. [DOI] [PubMed] [Google Scholar]
  19. Venta P. J., Shows T. B., Curtis P. J., Tashian R. E. Polymorphic gene for human carbonic anhydrase II: a molecular disease marker located on chromosome 8. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4437–4440. doi: 10.1073/pnas.80.14.4437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Whitney P. L., Briggle T. V. Membrane-associated carbonic anhydrase purified from bovine lung. J Biol Chem. 1982 Oct 25;257(20):12056–12059. [PubMed] [Google Scholar]

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