title: Purification of a mitochondrial DNA polymerase from Crithidia fasciculata.
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 A 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.
authors.affiliation: Department of Biological Chemistry, School of Medicine, Johns Hopkins University, Baltimore, Maryland 21205.
authors.forename: A F
authors.initials: AF
authors.lastname: Torri
authors.forename: P T
authors.initials: PT
authors.lastname: Englund
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002850:Chromatography, Gel; D016831:Crithidia fasciculata / Q000201:enzymology*; D003429:Cross Reactions; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011233:Precipitin Tests
chemical_list: D003854:Deoxyribonucleotides; D004259:DNA-Directed DNA Polymerase