Characterization of acrolein-induced protein cross-links.
Lipid peroxidation products contribute to protein aggregation that occurs during oxidative stress in a number of degenerative disorders. Acrolein (ACR), a highly toxic lipid peroxidation aldehyde, is a strong cross-linking agent of cellular components such as proteins. To understand the mechanisms of oxidative stress-induced protein aggregation, this study characterized the ACR modification of chain B from bovine insulin by mass spectrometry. To identify the cross-linking sites, the ACR-treated peptide was digested with a protease and the resulting peptides were analysed by liquid chromatography-tandem mass spectrometry. Inter- and intra-molecular cross-linking adducts were identified between amino groups and the side chain of histidine in the peptide. These results indicated that the ACR-induced cross-links were accompanied by two reactions, namely Michael addition and Schiff base formation. In conclusion, the use of mass spectrometric techniques provided chemical evidence for protein cross-linking with ACR.
Ishii
Takeshi
T
Department of Food and Nutritional Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan. ishii_t@u-shizuoka-ken.ac.jp
Yamada
Tomoe
T
Mori
Taiki
T
Kumazawa
Shigenori
S
Uchida
Koji
K
Nakayama
Tsutomu
T
D000171:Acrolein / Q000494:pharmacology*; D000595:Amino Acid Sequence; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D007328:Insulin / Q000737:chemistry; D015227:Lipid Peroxidation / Q000187:drug effects; D008954:Models, Biological; D010446:Peptide Fragments / Q000737:chemistry / Q000187:drug effects; D011506:Proteins / Q000737:chemistry / Q000187:drug effects*; D053719:Tandem Mass Spectrometry
D003432:Cross-Linking Reagents; D007328:Insulin; D010446:Peptide Fragments; D011506:Proteins; D000171:Acrolein; C041903:insulin B (9-30)