Leishmania tarentolae: a parallel isolation of cytochrome bc(1) and cytochrome c oxidase.
A rapid and simple method which allowed for a parallel isolation of cytochrome c reductase (cytochrome bc(1) ) and cytochrome c oxidase from kinetoplast-mitochondria of Leishmania tarentolae was developed. The method involved the lysis of kinetoplasts with dodecyl maltoside in the presence of 260 mM NaCl, followed by purification of bc(1) complexes on DEAE-sepharose CL-6B. The oxidase which was found in the flow-through fractions of the first chromatographic step was diluted and then repurified on a similar DEAE-sepharose column. The investigated properties of the isolated cytochrome c oxidase and reductase, such as their absolute and difference spectrum absorption maxima, heme content, specific activity, and subunit composition, confirm the usefulness of this method for obtaining highly active preparations of the enzymes.
Horváth
A
A
Department of Biology, University of California, Riverside, California 92521, USA.
Berry
E A
EA
Huang
L S
LS
Maslov
D A
DA
D000595:Amino Acid Sequence; D000818:Animals; D002852:Chromatography, Ion Exchange; D014450:Electron Transport Complex III / Q000737:chemistry / Q000302:isolation & purification*; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006418:Heme / Q000032:analysis; D007891:Leishmania / Q000201:enzymology*; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D013053:Spectrophotometry
D006418:Heme; D003576:Electron Transport Complex IV; D014450:Electron Transport Complex III