Table 4.
Comparison of lysine posttranslational modifications at individual sites of skin type I collagen from P3H3 and LH1 null mice relative to their wild-type controls
| Lys (%) | p value | Hyl (%) | p value | GHL (%) | p value | GGHL (%) | p value | |||
|---|---|---|---|---|---|---|---|---|---|---|
| α1 K87 | P3H3 | WT | 15.7 ± 13.3 | 9.78E-05 | 2.35 ± 0.24 | 1.01E-05 | 7.35 ± 1.08 | 1.03E-04 | 74.6 ± 12.3 | 4.49E-05 |
| Null | 77.8 ± 3.06 | 10.3 ± 1.15 | 2.30 ± 0.28 | 9.65 ± 1.78 | ||||||
| LH1 | WT | 8.28 ± 5.34 | 4.34E-08 | 2.06 ± 0.37 | 1.59E-04 | 6.83 ± 0.36 | 2.77E-08 | 82.8 ± 5.18 | 6.40E-08 | |
| Null | 98.9 ± 0.12 | 0.47 ± 0.78 | 0.20 ± 0.01 | 0.42 ± 0.03 | ||||||
| α2 K87 | P3H3 | WT | 6.41 ± 3.13 | 2.95E-08 | 93.6 ± 3.13 | 2.95E-08 | ND | ND | ||
| Null | 73.0 ± 1.92 | 27.0 ± 1.92 | ND | ND | ||||||
| LH1 | WT | 2.40 ± 0.28 | 3.33E-15 | 97.6 ± 0.28 | 3.33E-15 | ND | ND | |||
| Null | 98.0 ± 0.24 | 2.04 ± 0.24 | ND | ND | ||||||
| α1 K99 | P3H3 | WT | 76.5 ± 2.31 | 3.83E-05 | 20.4 ± 1.43 | 1.27E-05 | 2.86 ± 0.91 | 0.00916 | 0.32 ± 0.11 | 0.08426 |
| Null | 91.0 ± 1.40 | 7.68 ± 1.31 | 1.11 ± 0.12 | 0.19 ± 0.06 | ||||||
| LH1 | WT | 78.4 ± 0.82 | 1.21E-08 | 17.5 ± 1.06 | 1.46E-07 | 3.54 ± 0.23 | 4.29E-07 | 0.62 ± 0.05 | 9.04E-06 | |
| Null | 96.5 ± 0.26 | 2.70 ± 0.13 | 0.64 ± 0.11 | 0.18 ± 0.03 | ||||||
| α1 K174 | P3H3 | WT | 66.7 ± 4.73 | 3.38E-04 | 32.2 ± 4.34 | 2.82E-04 | 1.16 ± 0.44 | 0.00656 | ND | |
| Null | 84.2 ± 0.84 | 15.5 ± 0.79 | 0.24 ± 0.10 | ND | ||||||
| LH1 | WT | 63.1 ± 0.50 | 3.00E-11 | 34.4 ± 0.50 | 5.13E-11 | 1.88 ± 0.06 | 9.56E-09 | 0.53 ± 0.02 | ||
| Null | 92.1 ± 0.07 | 7.69 ± 0.11 | 0.24 ± 0.04 | ND | ||||||
| α2 K174 | P3H3 | WT | 40.6 ± 13.1 | 2.16E-04 | 4.86 ± 1.24 | 0.48365 | 50.6 ± 10.6 | 9.59E-05 | 3.94 ± 1.76 | |
| Null | 92.6 ± 1.27 | 5.40 ± 0.73 | 2.00 ± 1.21 | ND | ||||||
| LH1 | WT | 22.7 ± 1.17 | 4.31E-11 | 7.89 ± 0.80 | 3.12E-04 | 60.0 ± 0.79 | 9.10E-12 | 9.47 ± 0.48 | 2.07E-08 | |
| Null | 92.9 ± 0.58 | 4.59 ± 0.39 | 2.35 ± 0.23 | 0.20 ± 0.02 | ||||||
| α1 K219 | P3H3 | WT | 86.2 ± 1.55 | 0.00779 | 13.8 ± 1.55 | 0.00779 | ND | ND | ||
| Null | 90.0 ± 1.11 | 10.0 ± 1.11 | ND | ND | ||||||
| LH1 | WT | 85.3 ± 0.48 | 1.67E-07 | 14.7 ± 0.48 | 1.67E-07 | ND | ND | |||
| Null | 95.1 ± 0.54 | 4.91 ± 0.54 | ND | ND | ||||||
| α2 K219 | P3H3 | WT | 55.1 ± 7.05 | 0.00133 | 34.0 ± 4.12 | 0.0033 | 0.62 ± 0.21 | 0.00161 | 10.3 ± 2.77 | 5.15E-04 |
| Null | 75.2 ± 0.91 | 23.9 ± 1.09 | 0.03 ± 0.04 | 0.88 ± 0.25 | ||||||
| LH1 | WT | 50.3 ± 0.79 | 6.51E-10 | 34.7 ± 0.92 | 2.31E-08 | 0.91 ± 0.13 | 1.70E-05 | 13.9 ± 0.63 | 1.42E-08 | |
| Null | 85.4 ± 0.65 | 13.7 ± 0.64 | 0.08 ± 0.02 | 0.80 ± 0.09 | ||||||
| α1 K564 | P3H3 | WT | 77.3 ± 1.20 | 0.7026 | 20.3 ± 1.06 | 0.33603 | 1.72 ± 0.13 | 0.07732 | 0.70 ± 0.12 | 0.13392 |
| Null | 77.6 ± 0.79 | 19.7 ± 0.65 | 1.93 ± 0.14 | 0.85 ± 0.13 | ||||||
| LH1 | WT | 78.9 ± 0.82 | 1.20E-07 | 18.4 ± 0.58 | 4.12E-08 | 2.00 ± 0.19 | 7.87E-06 | 0.69 ± 0.10 | 3.30E-04 | |
| Null | 92.4 ± 0.47 | 6.76 ± 0.35 | 0.55 ± 0.07 | 0.28 ± 0.05 | ||||||
| α2 K933 | P3H3 | WT | ND | 93.1 ± 0.37 | 2.12E-09 | 6.86 ± 2.37 | ND | |||
| Null | 86.7 ± 1.56 | 13.3 ± 1.56 | ND | ND | ||||||
| LH1 | WT | ND | 93.0 ± 1.23 | 7.36E-12 | 7.02 ± 1.23 | ND | ||||
| Null | 97.9 ± 0.28 | 2.05 ± 0.28 | ND | ND | ||||||
| Lys-Lys (%) | p value | Lys-Hyl (%) | p value | Hyl-Hyl (%) | p value | |||
|---|---|---|---|---|---|---|---|---|
| α1 K918/K930 | P3H3 | WT | 1.57 ± 0.73 | 5.07E-07 | 8.11 ± 2.66 | 0.09689 | 90.3 ± 3.17 | 3.67E-07 |
| Null | 88.9 ± 7.73 | 5.16 ± 1.41 | 5.99 ± 6.36 | |||||
| LH1 | WT | 0.29 ± 0.21 | 1.11E-16 | 4.00 ± 0.18 | 4.47E-08 | 95.7 ± 0.21 | 3.33E-16 | |
| Null | 99.0 ± 0.06 | 0.78 ± 0.07 | 0.22 ± 0.12 | |||||
GGHL, glucosylgalactosyl hydroxylysine; GHL, galactosyl hydroxylysine; Hyl, unmodified hydroxylysine; Hyl-Hyl, unmodified hydroxylysine and unmodified hydroxylysine; Lys, unmodified lysine; Lys-Hyl, unmodified lysine and unmodified hydroxylysine; Lys-Lys, unmodified lysine and unmodified lysine.
Values are given as means ± S.D. Biological replicates were n = 4 for all tissues and genotypes.
The lysine modification sites α1 K918 and K930 were included in the same peptide GDKGETGEQGDRGIKGER.
Lys + Hyl + GHL + GGHL = 100% and Lys-Lys + Lys-Hyl + Hyl-Hyl = 100%. Values of modified and unmodified hydroxylysine and unmodified lysine were obtained using mass spectrometry.