A Role for Iron in an Ancient Carbonic Anhydrase

Brian C. Tripp, Caleb B. Bell, Francisco Cruz, Carsten Krebs, James Gregory Ferry

Research output: Contribution to journalArticle

109 Citations (Scopus)

Abstract

Since 1933, carbonic anhydrase research has focused on enzymes from mammals (α class) and plants (β class); however, two additional classes (γ and δ) were discovered recently. Cam, from the procaryote Methanosarcina thermophila, is the prototype of the γ class and the first carbonic anhydrase to be characterized from either an anaerobic organism or the Archaea domain. All of the enzymes characterized from the four classes have been purified aerobically and are reported to contain a catalytic zinc. Herein, we report the anaerobic reconstitution of apo-Cam with Fe 2+, which yielded Cam with an effective kcat that exceeded that for the Zn2+-reconstituted enzyme. Mössbauer spectroscopy showed that the Fe2+-reconstituted enzyme contained high spin Fe2+ that, when oxidized to Fe3+, inactivated the enzyme. Reconstitution with Fe3+ was unsuccessful. Reconstitution with Cu2+, Mn2+, Ni2+, or Cd2+ yielded enzymes with effective kcat values that were 10% or less than the value for the Zn2+-reconstituted Cam. Cam produced in Escherichia coli and purified anaerobically contained iron with effective kcat and kcat/Km values exceeding the values for Zn2+-reconstituted Cam. The results identify a previously unrecognized biological function for iron.

Original languageEnglish (US)
Pages (from-to)6683-6687
Number of pages5
JournalJournal of Biological Chemistry
Volume279
Issue number8
DOIs
StatePublished - Feb 20 2004

Fingerprint

Carbonic Anhydrases
Cams
Iron
Enzymes
Methanosarcina
Mammals
Archaea
Escherichia coli
Zinc
Spectrum Analysis
Spectroscopy
Research

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Tripp, Brian C. ; Bell, Caleb B. ; Cruz, Francisco ; Krebs, Carsten ; Ferry, James Gregory. / A Role for Iron in an Ancient Carbonic Anhydrase. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 8. pp. 6683-6687.
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A Role for Iron in an Ancient Carbonic Anhydrase. / Tripp, Brian C.; Bell, Caleb B.; Cruz, Francisco; Krebs, Carsten; Ferry, James Gregory.

In: Journal of Biological Chemistry, Vol. 279, No. 8, 20.02.2004, p. 6683-6687.

Research output: Contribution to journalArticle

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