Detection and characterization of in vivo nitration and oxidation of tryptophan residues in proteins.
Oxygen and nitrogen centered reactive species can cause specific structural modifications in amino acids and proteins, such as the addition of a nitro group onto aromatic residues. Heretofore, studies on protein nitration have mainly focused on the in vitro and in vivo nitro addition to tyrosine residues (3-nitrotyrosine or 3NT), whereas the formation of nitrotryptophan in proteins in vivo and/or its functional significance has remained quite obscure. A novel structural modification, involving the addition of nitro and hydroxy groups to tryptophan, has been detected in the mitochondrial protein succinyl-CoA:3-oxoacid CoA transferase (SCOT) in rat heart. Modified SCOT accumulated progressively with age, which was associated with an elevation of its activity. The specific biochemical properties of this novel amino acid were characterized by a combination of HPLC-electrochemical detection and mass spectrometric analysis. This chapter describes the experimental steps involved in the characterizations and a procedure for the synthesis of nitrohydroxytryptophan. Similar methodology can be applied to the identification of nitrohydroxytryptophan in other proteins.
Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA, USA.
Catherine
C
Bregere
Igor
I
Rebrin
Rajindar S
RS
Sohal
D000818:Animals; D006801:Humans; D009566:Nitrates / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D014364:Tryptophan / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
D009566:Nitrates; D011506:Proteins; D014364:Tryptophan