Purification of a mitochondrial DNA polymerase from Crithidia fasciculata

Al F. Torri, Paul T. Englund

Research output: Contribution to journalArticle

Abstract

The mitochondrial DNA polymerase from Crithidia fasciculata has been purified to near homogeneity. SDS-PAGE analysis of the purified enzyme reveals a single polypeptide with a molecular weight of approximately 43,000. The protein is basic, with an isoelectric point between 7.6-8.0. Its Stokes radius of 22 Å and its sedimentation coefficient of 4.1 S suggest a native molecular weight of 38,000, indicating that the protein is a monomer under our experimental conditions. Western blots and immunoprecipitations of crude extracts reveal a cross-reacting protein of 48 kDa, suggesting that the purified enzyme may be an enzymatically active proteolytic product. The mitochondrial origin of the polymerase was confirmed by cell fractionation. Our results indicate that the C. fasciculata enzyme may be among the smallest known mitochondrial polymerases.

Original languageEnglish (US)
Pages (from-to)4786-4792
Number of pages7
JournalJournal of Biological Chemistry
Volume267
Issue number7
StatePublished - Mar 5 1992

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Crithidia fasciculata
DNA-Directed DNA Polymerase
Mitochondrial DNA
Purification
Enzymes
Molecular Weight
Molecular weight
Cell Fractionation
Proteins
Isoelectric Point
Fractionation
Complex Mixtures
Sedimentation
Immunoprecipitation
Polyacrylamide Gel Electrophoresis
Monomers
Western Blotting
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification of a mitochondrial DNA polymerase from Crithidia fasciculata. / Torri, Al F.; Englund, Paul T.

In: Journal of Biological Chemistry, Vol. 267, No. 7, 05.03.1992, p. 4786-4792.

Research output: Contribution to journalArticle

Torri, Al F. ; Englund, Paul T. / Purification of a mitochondrial DNA polymerase from Crithidia fasciculata. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 7. pp. 4786-4792.
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