Induction of apoptotic program in cell-free extracts : requirement for datp and cytochrome c

A thermodynamic approach to the problem of stabilization of globular protein structure: A calorimetric study

Correlation of the kinetics of electron transfer activity of various eukaryotic cytochromes c with binding to mitochondrial cytochrome c oxidase.

Definition of cytochrome c binding domains by chemical modification. III. Kinetics of reaction of carboxydinitrophenyl cytochromes c with cytochrome c oxidase.

Design, synthesis and biological evaluation of some pyrazole derivatives as anti-inflammatory-antimicrobial agents

Publisher Summary This chapter describes the preparation procedure, purification, and properties of the cytochrome c from vertebrate and invertebrate sources. Cytochrome c is extracted from ground and homogenized tissue with a dilute solution of aluminum sulfate at pH 4.5. At slightly alkaline pH, excess aluminum ions are precipitated as the hydroxide, and exchanged for three monovalent ammonium ions. This permits the direct collection of the protein from the extract on a cation exchange resin. The cytochrome c is purified by (NH 4 ) 2 SO 4 fractionation, cation exchange chromatography, and crystallization. Other methods, involving extraction with trichloroacetic or sulfuric acids, extraction at neutral pH following the decomposition of the tissue in acetic acid or by treatment with a suerose-saponin solution and acetone are described. Cytochrome c is a basic protein with a molecular weight near 12,300. It consists of single polypeptide chain 103-108 amino acid residues long, to which a single heme prosthetic group is covalently attached by thioether bonds formed by the addition of the sulfhydryl groups of two cysteinyl residues in positions 14 and 17 across the vinyl side chains of the porphyrin ring. In the vertebrate cytochromes c the amino-terminal residue is acetylglyeine, while the proteins from nonvertebrate sources carry several extra residues in place of the acetyl.

E. Margoliash

Papers

[61] Cytochrome c from vertebrate and invertebrate sources