TABLE 1

Glycation adducts in human type IV collagen minimally modified by methylglyoxal, glucose, hyperglycemic, and normoglycemic medium

Glycation adductControl-1*MGmin-collagen (6-h incubation)MG-collagen (12-h incubation)MG-collagen (24-h incubation)Control-2AGEmin-collagen§Conditioned collagen (5 mmol/l glucose)Conditioned collagen (20 mmol/l glucose)
MG-H196.9 ± 9.53,600 ± 1657,553 ± 46010,506 ± 614136 ± 62,667 ± 60333 ± 28611 ± 91
Argpyrimidine7.1 ± 1.051.7 ± 18.8106.5 ± 26.9224.0 ± 39.35.0 ± 1.569.8 ± 12.547.9 ± 3.3101.6 ± 4.2
CEL12.0 ± 1.029.0 ± 7.324.0 ± 4.0220.9 ± 10.823.8 ± 4.2102.6 ± 22.432.3 ± 1.164.5 ± 16.7
MOLD1.4 ± 0.31.5 ± 0.14.5 ± 1.19.8 ± 1.44.5 ± 1.4148.2 ± 28.412.5 ± 6.7118.9 ± 32.7
CMA8.6 ± 0.915.5 ± 1.829.8 ± 9.522.0 ± 3.76.9 ± 3.4888.9 ± 87.315.9 ± 3.219.4 ± 3.9
Fructosyl-lysine104.8 ± 7.2104.7 ± 13.6169.2 ± 44.2149.1 ± 39.173.1 ± 42.21,909.2 ± 233.9119.0 ± 27.7126.6 ± 53.3
G-H14.0 ± 0.86.8 ± 2.73.9 ± 0.94.7 ± 1.515.1 ± 1.91,134.6 ± 346.122.8 ± 9.221.1 ± 4.4
3DG-H45.7 ± 10.156.1 ± 4.446.1 ± 6.140.5 ± 16.219.5 ± 0.875.7 ± 23.029.0 ± 9.333.3 ± 1.7
CML36.0 ± 4.040.0 ± 3.064.0 ± 7.057.0 ± 14.047.7 ± 4.3454.8 ± 37.193.1 ± 3.894.3 ± 3.3
Nitrotyrosine8.1 ± 1.910.3 ± 1.67.2 ± 1.36.9 ± 3.33.4 ± 0.218.2 ± 7.116.5 ± 0.913.6 ± 0.4
Pentosidine0.43 ± 0.070.33 ± 0.030.30 ± 0.050.32 ± 0.050.79 ± 0.2314.4 ± 1.20.38 ± 0.050.40 ± 0.05
Dityrosine0.19 ± 0.060.45 ± 0.040.47 ± 0.090.50 ± 0.030.51 ± 0.131.25 ± 0.310.40 ± 0.040.45 ± 0.09
  • Data are mean ± SD (n = 3) (mmol/mol triple-chain collagen). Incubation conditions were as follows.

  • *

    * Collagen (6 mg/ml) in 100 mmol/l sodium phosphate buffer, pH 7.4, at 37oC, for 24 h.

  • Collagen (6 mg/ml) with 500 μmol/l methylglyoxal in 100 mmol/l sodium phosphate buffer, pH 7.4, at 37oC, for the times indicated.

  • Collagen (6 mg/ml) in 100 mmol/l sodium phosphate buffer, pH 7.4, at 37oC, for 21 days.

  • §

    § Collagen with 50 mmol/l β-d-glucose in 100 mmol/l sodium phosphate buffer, pH 7.4, at 37oC, for 21 days.

  • Collagen (6 mg/ml) with medium from endothelial cells cultured for 7 days with the concentration of glucose indicated, pH 7.4, at 37oC, for 24 h. 3DG-H, Nδ-(5-hydro-5-(2,3,4-trihydroxybutyl)-4-imidazolon-2-yl)orinithine and related structural isomers; CEL, Nε-(1-carboxyethyl)lysine; CMA, Nω-carboxymethylarginine; CML, Nε-carboxymethyllysine; G-H1, Nδ-(5-hydro-4-imidazolon-2-yl)ornithine; MOLD, methylglyoxal-derived lysine dimer.