Fructosamine 3-kinase is involved in an intracellular deglycation pathway in human erythrocytes

Biochem J. 2002 Aug 1;365(Pt 3):801-8. doi: 10.1042/BJ20020325.

Abstract

Fructosamine 3-kinase, which phosphorylates low-molecular-mass and protein-bound fructosamines on the third carbon of their deoxyfructose moiety, is quite active in erythrocytes, and was proposed to initiate a process removing fructosamine residues from proteins. In the present study, we show that incubation of human erythrocytes with 200 mM glucose not only caused the progressive formation of glycated haemoglobin, but also increased the level of an anionic form of haemoglobin containing alkali-labile phosphate, to approx. 5% of total haemoglobin. 1-Deoxy-1-morpholinofructose (DMF), a substrate and competitive inhibitor of fructosamine 3-kinase, doubled the rate of accumulation of glycated haemoglobin, but markedly decreased the amount of haemoglobin containing alkali-labile phosphate. The latter corresponds therefore to haemoglobin bound to a fructosamine 3-phosphate group (FN3P-Hb). Returning erythrocytes incubated with 200 mM glucose and DMF to a low-glucose medium devoid of DMF caused a decrease in the amount of glycated haemoglobin, a transient increase in FN3P-Hb and a net decrease in the sum (glycated haemoglobin+FN3P-Hb). These effects were prevented by DMF, indicating that fructosamine 3-kinase is involved in the removal of fructosamine residues. The second step of this 'deglycation' process is most likely a spontaneous decomposition of the fructosamine 3-phosphate residues to a free amine, 3-deoxyglucosone and P(i). This is consistent with the findings that 2-oxo-3-deoxygluconate, the product of 3-deoxyglucosone oxidation, is formed in erythrocytes incubated for 2 days with 200 mM glucose in a sufficient amount to account for the removal of fructosamine residues from proteins, and that DMF appears to inhibit the formation of 2-oxo-3-deoxygluconate from elevated glucose concentrations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Erythrocytes / enzymology*
  • Fructose / analogs & derivatives
  • Fructose / metabolism
  • Glucose / metabolism*
  • Glycated Hemoglobin A / metabolism*
  • Humans
  • Lysine / analogs & derivatives*
  • Lysine / metabolism
  • Molecular Structure
  • Morpholines / metabolism
  • Phosphorylation
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism*
  • Recombinant Proteins / metabolism

Substances

  • Glycated Hemoglobin A
  • Morpholines
  • Recombinant Proteins
  • fructosyl-lysine
  • Fructose
  • 1-deoxy-1-morpholinofructose
  • Phosphotransferases (Alcohol Group Acceptor)
  • fructosamine-3-kinase
  • Glucose
  • Lysine