Development of a derivatisation method for the analysis of aldehyde modified amino acid residues in proteins by Fourier transform mass spectrometry.
A method was developed for the analysis of amino acids within bovine serum albumin (BSA) which had been modified by reaction with different enals. BSA was reacted with the aldehydes and the reaction products were stabilised by reaction with NaBH(4). The protein was then hydrolysed with 6N HCl and the hydrolysis products were analysed by liquid chromatography-mass spectrometry (LC-MS). The modified amino acids were derivatised with propylchloroformate. High resolution mass spectrometry carried out using an LTQ-Orbitrap instrument which was able to characterise a wide range of adducts. In addition double adducts were observed to be formed with 4-hydroxynonenal (HNE) and lysine or lysine+histidine. Qualitatively it was possible to consistently observe a pyridinium adduct formed between lysine and pentenal in human plasma from normal subjects.
Pournamdari
Mostafa
M
Strathclyde Institute for Pharmacy and Biomedical Science (SIPBS), 27 Taylor Street, Glasgow G4 0NR, United Kingdom.
Saadi
Ahmed
A
Ellis
Elizabeth
E
Andrew
Ruth
R
Walker
Brian
B
Watson
David G
DG
D000447:Aldehydes / Q000032:analysis* / Q000737:chemistry; D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D001798:Blood Proteins / Q000032:analysis*; D002853:Chromatography, Liquid; D005583:Fourier Analysis; D006639:Histidine / Q000737:chemistry; D006801:Humans; D008239:Lysine / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D010880:Piperidines / Q000737:chemistry; D011726:Pyridinium Compounds / Q000737:chemistry; D012545:Schiff Bases / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000032:analysis*
D000447:Aldehydes; D000596:Amino Acids; D001798:Blood Proteins; D010880:Piperidines; D011726:Pyridinium Compounds; D012545:Schiff Bases; D012710:Serum Albumin, Bovine; D006639:Histidine; C027576:4-hydroxy-2-nonenal; D008239:Lysine