Glutathione adducts, not carbamylated lysines, are the major modification of lens alpha-crystallins from renal failure patients.
alpha-Crystallins from the water-soluble and the water-insoluble, guanidine-soluble portions of lenses from four renal failure patients and two normal donors of similar age were isolated and enzymatically digested into peptides. Molecular weights of the peptides, determined by fast atom bombardment mass spectrometry, indicated modifications specifically associated with renal failure. The only modifications observed in the alpha-crystallins from renal failure patients, but not in the normal old lenses, were glutathione adducts to Cys 131 and Cys 142. These adducts were present in the lenses of all four renal failure patients, but not in the two normal old lenses. The four lenses from the renal failure patients were searched for evidence of carbamylation at lysyl or cysteinyl residues: carbamylation was not detected. Because the same mass spectrometric methods had previously demonstrated sufficient sensitivity and specificity to detect as little as 5% modification in the examination of in vitro carbamylated bovine lenses, these results indicated that carbamylation is not a major modification of the lens alpha-crystallins of renal failure patients.
Smith
J B
JB
Department of Medicinal Chemistry and Pharmacognosy, Purdue University, West Lafayette, Indiana 47907, USA.
Shun-Shin
G A
GA
Sun
Y
Y
Miesbauer
L R
LR
Yang
Z
Z
Yang
Z
Z
Zhou
X
X
Schwedler
J
J
Smith
D L
DL
D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000378:metabolism; D002219:Carbamates / Q000378:metabolism*; D003459:Crystallins / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D005978:Glutathione / Q000378:metabolism*; D019791:Guanidine; D006146:Guanidines / Q000737:chemistry; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D051437:Renal Insufficiency / Q000378:metabolism*; D012995:Solubility; D016339:Spectrometry, Mass, Fast Atom Bombardment; D014867:Water / Q000737:chemistry
D002219:Carbamates; D003459:Crystallins; D006146:Guanidines; D014867:Water; D005978:Glutathione; D019791:Guanidine; D008239:Lysine