Modification of horse heart cytochrome c with trans-2-hexenal.
Horse heart cytochrome c reacting with trans-2-hexenal was used as a simple model of the nonspecific interactions of proteins with 2-alkenals. The reaction mixtures containing relatively high concentrations of the protein and aldehyde were characterized using visible spectrophotometry, fluorescence, and circular dichroism measurement, capillary isoelectric focusing, size-exclusion chromatography, polyacrylamide gel electrophoresis, and mass-spectrometric techniques. The mass-spectrometric data indicate that cytochrome c becomes modified with one or two molecules of hexenal as the major reaction product. The modified species with a correspondingly lowered isoelectric point were detected through capillary isoelectric focusing. The results of proteolytic studies indicate nonspecific modifications. Significant quantities of the oligomeric forms of hexenal-modified protein were also observed electrophoretically.
Zídek
L
L
Department of Chemistry, Indiana University, Bloomington 47405-4001, USA.
Dolezel
P
P
Chmelík
J
J
Baker
A G
AG
Novotny
M
M
D000447:Aldehydes / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D003574:Cytochrome c Group / Q000737:chemistry / Q000187:drug effects*; D006736:Horses; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology*; D010455:Peptides / Q000737:chemistry
D000447:Aldehydes; D003574:Cytochrome c Group; D010455:Peptides; C051750:2-hexenal