The reaction of bovine lens alpha A-crystallin with aspirin.
Acetylation of the lysines of bovine lens alpha A-crystallin has been examined after 0-48-hr incubations of whole alpha-crystallins in 100 mM aspirin. The alpha A-crystallins were isolated after the incubation, proteolytically digested into peptides which were separated by reversed phase HPLC and analysed by mass spectrometry. For the reaction conditions used in this investigation, acetylated lysyl residues were the principal products. The extent of acetylation was quantified from the intensities of the peaks in the fast atom bombardment mass spectra of the modified and unmodified peptides. The modified lysine containing peptides demonstrated that all seven lysyl residues of alpha A-crystallin reacted with aspirin; the extent of acetylation at each lysyl residue varied. Plots of the extent of acetylation vs. time were used to calculate rate constants for the reaction at each lysyl residue. The rate constant for the acetylation of Lys 166, the most reactive, was about seven times greater than for Lys 88, the least reactive. These rate constants were used to calculate the yield of predicted products for the reaction of alpha-crystallin with therapeutic concentrations of aspirin. Comparison of the yield of acetylated alpha-crystallin with the yield of carbamylated alpha-crystallin that might occur due to renal failure indicates that aspirin is not likely to be an effective inhibitor of cataract due to carbamylation of lysyl residues.
Hasan
A
A
H.E.J. Research Institute of Chemistry, University of Karachi, Pakistan.
Smith
J B
JB
Qin
W
W
Smith
D L
DL
D000107:Acetylation; D000595:Amino Acid Sequence; D000818:Animals; D001241:Aspirin / Q000494:pharmacology*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003459:Crystallins / Q000187:drug effects*; D066298:In Vitro Techniques; D007700:Kinetics; D008239:Lysine / Q000032:analysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D013997:Time Factors
D003459:Crystallins; D008239:Lysine; D001241:Aspirin