Particulate carbonic anhydrase in homogenates of human kidney

Dana N. McKinley, Philip L. Whitney

Research output: Contribution to journalArticle

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Abstract

About 2% of human kidney carbonic anhydrase (carbonate hydro-lyase, EC 4.2.1.1) has been found in particulate fractions. Its distribution in the particulate fractions obtained by differential centrifugation suggests that it may be concentrated in the brush border. The particulate enzyme is like red cell carbonic anhydrace C in its susceptibility to inhibition by anions. Particulate carbonic anhydrase is firmly bound to the membrane and is not released by incubation at pH 10.6 and 37°C or by addition of Triton X-100 or deoxycholate. In 10% Triton X-100 at pH 11.3 and 37°C, the particulate enzyme is inactivated with a half time of about 20 min, and this is at least an order of magnitude slower than the inactivation of soluble enzymes in the presence or absence of membranes. The soluble enzymes are inactivated within a few minutes at 25°C in 3-4% sodium dodecyl sulfate, but the particulate enzyme is relatively stable under those conditions, and its half-time of inactivation at 14°C with a detergent-protein ratio of 25 was about 24 h. Gel filtration with Ultragel AcA-44 in sodium dodecyl sulfate indicates that the membrane carbonic anhydrase has a molecular weight of less than 66 000, so its stability is not due to association with large membrane fragments or vesicles. These results suggest that the membrane enzyme may be a different isozyme than the soluble carbonic anhydrases. Although present in relatively small amounts, its localization on the membrane could give it functional significance.

Original languageEnglish
Pages (from-to)780-790
Number of pages11
JournalBBA - Enzymology
Volume445
Issue number3
DOIs
StatePublished - Oct 11 1976
Externally publishedYes

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Carbonic Anhydrases
Kidney
Membranes
Enzymes
Octoxynol
Sodium Dodecyl Sulfate
Deoxycholic Acid
Microvilli
Centrifugation
Detergents
Isoenzymes
Gel Chromatography
Anions
Molecular Weight
Proteins

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Particulate carbonic anhydrase in homogenates of human kidney. / McKinley, Dana N.; Whitney, Philip L.

In: BBA - Enzymology, Vol. 445, No. 3, 11.10.1976, p. 780-790.

Research output: Contribution to journalArticle

McKinley, Dana N. ; Whitney, Philip L. / Particulate carbonic anhydrase in homogenates of human kidney. In: BBA - Enzymology. 1976 ; Vol. 445, No. 3. pp. 780-790.
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