Oxidized amino acids in lens protein with age. Measurement of o-tyrosine and dityrosine in the aging human lens.
The concentrations of ortho-tyrosine (o-Tyr) and dityrosine (DT) were measured in noncataractous human lenses in order to assess the role of protein oxidation reactions in the aging of lens proteins. The measurements were conducted by selected ion monitoring-gas chromatography/mass spectrometry using deuterium-labeled internal standards, which provided both high sensitivity and specificity for the quantitation of o-Tyr and DT. Between ages 1 and 78 years, the o-Tyr concentration in lens proteins varied from 0.3 to 0.9 mmol of o-Tyr/mol of Phe (n = 19), while DT ranged from 1 to 3 mumol of DT/mol of Tyr (n = 30). There were no significant changes in levels of o-Tyr with lens age. There was a statistically significant, but only slight, increase in DT in lens proteins with age (approximately 33% increases between ages 1 and 78, r = 0.5, p < 0.01). At the same time, total protein fluorescence, measured at DT wavelengths (Ex = 317 nm, Em = 407 nm), increased 11-fold between ages 1 and 78 and correlated strongly with age (r = 0.82, p < 0.0001). Although the fluorescence maxima of lens proteins were similar to those of DT, DT accounted for less than 1% of the DT-like fluorescence in lens protein at all ages. These observations indicate that oxidation of Phe and Tyr plays a limited role in the normal aging of lens proteins in vivo.
Wells-Knecht
M C
MC
Department of Chemistry and Biochemistry, University of South Carolina, Columbia 29208.
Huggins
T G
TG
Dyer
D G
DG
Thorpe
S R
SR
Baynes
J W
JW
D000293:Adolescent; D000328:Adult; D000368:Aged; D000375:Aging / Q000502:physiology*; D002648:Child; D002675:Child, Preschool; D003459:Crystallins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007223:Infant; D007536:Isomerism; D007908:Lens, Crystalline / Q000254:growth & development / Q000378:metabolism*; D008875:Middle Aged; D010084:Oxidation-Reduction; D013050:Spectrometry, Fluorescence; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis*
D003459:Crystallins; D014443:Tyrosine; C026741:2-tyrosine; C007543:dityrosine