title: Plasma proteins containing damaged L-isoaspartyl residues are increased in uremia: implications for mechanism.
abstract: BACKGROUND
  Several alterations of protein structure and function have been reported in uremia. Impairment of a transmethylation-dependent protein repair mechanism possibly related to a derangement in homocysteine metabolism is also present in this condition, causing erythrocyte membrane protein damage. Homocysteine may affect proteins via the accumulation of its parent compound S-adenosylhomocysteine (AdoHcy), a powerful in vivo methyltransferase inhibitor. However, since plasma homocysteine is mostly protein bound, a direct influence on protein structures cannot be ruled out. We measured the levels of L-isoaspartyl residues in plasma proteins of uremic patients on hemodialysis. These damaged residues are markers of molecular age, which accumulate when transmethylation-dependent protein repair is inhibited and/or protein instability is increased.
  
  
  METHODS
  L-isoaspartyl residues in plasma proteins were quantitated using human recombinant protein carboxyl methyl transferase (PCMT). Plasma concentrations of homocysteine metabolites were also measured under different experimental conditions in hemodialysis patients.
  
  
  RESULTS
  The concentration of damaged plasma proteins was increased almost twofold compared to control (controls 147.83 +/- 17.75, uremics 282.80 +/- 26.40 pmol of incorporated methyl groups/mg protein, P < 0.003). The major protein involved comigrated with serum albumin. Although hyperhomocysteinemia caused a redistribution of thiols bound to plasma proteins, this mechanism did not significantly contribute to the increase in isoaspartyl residues. The S-adenosylmethionine (AdoMet)/AdoHcy concentration ratio, an indicator of the flux of methyl group transfer, was altered. This ratio was partially corrected by folate treatment (0.385 +/- 0.046 vs. 0.682 +/- 0.115, P < 0.01), but protein L-isoaspartate content was not.
  
  
  CONCLUSIONS
  Plasma protein damage, as determined by protein L-isoaspartyl content, is increased in uremia. This alteration is to be ascribed to an increased protein structural instability, rather than the effect of hyperhomocysteinemia.
authors.affiliation: Division of Nephrology, Department of Pediatrics, Second University of Naples, School of Medicine and Surgery, Naples, Italy. alessandra.perna@unina2.it
authors.forename: A F
authors.initials: AF
authors.lastname: Perna
authors.forename: P
authors.initials: P
authors.lastname: Castaldo
authors.forename: N G
authors.initials: NG
authors.lastname: De Santo
authors.forename: E
authors.initials: E
authors.lastname: di Carlo
authors.forename: A
authors.initials: A
authors.lastname: Cimmino
authors.forename: P
authors.initials: P
authors.lastname: Galletti
authors.forename: V
authors.initials: V
authors.lastname: Zappia
authors.forename: D
authors.initials: D
authors.lastname: Ingrosso
mesh_terms: D001224:Aspartic Acid / Q000032:analysis / Q000097:blood*; D015415:Biomarkers; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005492:Folic Acid / Q000008:administration & dosage; D006397:Hematinics / Q000008:administration & dosage; D006710:Homocysteine / Q000097:blood*; D006801:Humans; D020138:Hyperhomocysteinemia / Q000378:metabolism; D008745:Methylation; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase; D011496:Protein Methyltransferases; D006435:Renal Dialysis; D012435:S-Adenosylhomocysteine / Q000097:blood*; D012709:Serum Albumin / Q000378:metabolism; D014511:Uremia / Q000097:blood* / Q000188:drug therapy
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D006397:Hematinics; D012709:Serum Albumin; D006710:Homocysteine; D001224:Aspartic Acid; D005492:Folic Acid; D012435:S-Adenosylhomocysteine; D011496:Protein Methyltransferases; D026601:Protein D-Aspartate-L-Isoaspartate Methyltransferase