Plasma protein aspartyl damage is increased in hemodialysis patients: studies on causes and consequences.
Plasma proteins in hemodialysis patients display a significant increase in deamidated/isomerized Asx (asparagine and aspartic acid) content, a marker of protein fatigue damage. This has been linked to the toxic effects of hyperhomocysteinemia in uremic erythrocytes; however, treatment aimed at abating homocysteine levels did not lead to significant reductions in plasma protein damage. The hypothesis that lack of reduction in protein damage could be due to protein increased intrinsic instability, as result of interference with the uremic milieu rather than to hyperhomocysteinemia, was put forward. The deamidated/isomerized Asx content of normal plasma incubated with several uremic toxins for 24 h, 72 h, and 7 d was measured, identifying a group of toxins that were able to elicit this kind of damage. Uremic toxins were also incubated with purified human albumin, and dose-response experiments with the two most toxic agents in terms of protein damage (guanidine and guanidinopropionic acid) were carried out. The effect of the hemodialysis procedure on protein damage was evaluated. For investigating also the consequences of these alterations, human albumin was treated in vitro to produce an increase in deamidated/isomerized Asx residues, and the effects of albumin deamidation on protein binding were evaluated. Among the uremic toxins that are able to elicit protein damage, guanidine produced a dose-dependent increase in protein damage. No difference was found after a hemodialysis session. Deamidated albumin shows normal binding capacity to warfarin, salicylic acid, or diazepam but reduced binding to homocysteine. In conclusion, uremic toxins, especially guanidine, display an ability to induce significant protein damage, which can in turn have functional consequences.
Perna
Alessandra F
AF
First Division of Nephrology, Second University of Naples, School of Medicine, Italy. alessandra.perna@unina2.it
Ingrosso
Diego
D
Satta
Ersilia
E
Lombardi
Cinzia
C
Galletti
Patrizia
P
D'Aniello
Antimo
A
De Santo
Natale G
NG
D001216:Asparagine / Q000032:analysis / Q000097:blood* / Q000187:drug effects; D001224:Aspartic Acid / Q000032:analysis / Q000097:blood* / Q000187:drug effects; D015415:Biomarkers / Q000097:blood; D001774:Blood Chemical Analysis; D001798:Blood Proteins / Q000032:analysis / Q000378:metabolism*; D003975:Diazepam / Q000494:pharmacology; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007676:Kidney Failure, Chronic / Q000097:blood* / Q000628:therapy; D008297:Male; D011379:Prognosis; D011485:Protein Binding; D006435:Renal Dialysis / Q000009:adverse effects* / Q000379:methods; D018570:Risk Assessment; D012459:Salicylates / Q000494:pharmacology; D012494:Sampling Studies; D012680:Sensitivity and Specificity; D014859:Warfarin / Q000494:pharmacology
D015415:Biomarkers; D001798:Blood Proteins; D012459:Salicylates; D001224:Aspartic Acid; D014859:Warfarin; D001216:Asparagine; D003975:Diazepam