Carbamoylation of hemoglobin in uremic patients determined by antibody specific for homocitrulline (carbamoylated epsilon-N-lysine).
Polyclonal antisera to epsilon-amino-carbamoyl-lysine (homocitrulline) on guinea pig carbamoylated low density lipoprotein were used as probes to identify homocitrulline. These antisera detect homocitrulline in the immunogen and on other carbamoylated blood proteins including those from different species. In vivo carbamoylation of intracellular hemoglobin obtained from uremic patients and carbamoyl-proteins, carbamoylated in vitro, located on erythrocyte and lymphocyte cell surface membranes were detected. Solid phase, direct and competitive ELISAs for homocitrulline were used to analyze carbamoylated blood proteins. In uremic patients carbamoylation of protein by endogenous urea-derived cyanate occurs on the epsilon-amino group of lysine residues. Identification of carbamoylated proteins from uremic patients using an immunochemical probe of site specific antibody provides a way to monitor post-translational modification of proteins due to uremia and may give further insight for understanding the pathophysiology of end stage renal disease.
Kraus
L M
LM
Department of Biochemistry, University of Tennessee, Memphis 38163.
Miyamura
S
S
Pecha
B R
BR
Kraus
A P
AP
D000818:Animals; D000915:Antibody Affinity; D002219:Carbamates; D002956:Citrulline / Q000031:analogs & derivatives* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D006455:Hemoglobins, Abnormal / Q000032:analysis*; D007676:Kidney Failure, Chronic / Q000097:blood*; D008077:Lipoproteins, LDL / Q000276:immunology; D014511:Uremia / Q000097:blood*
D002219:Carbamates; D006455:Hemoglobins, Abnormal; D008077:Lipoproteins, LDL; C001352:homocitrulline; D002956:Citrulline