BSDB

The conformational change and the identification of the mechanical clamp for proteins can be derived by looking at the unfolding scenario. A unfolding scenario shows distance d_u at which a given native contact is broken for the last time. Contacts in the scenario diagrams are identified by the sequential distance |j - i|. A contact is said to be broken if the distance between amino acids i and j exceeds 1.5σ_ij. An accumulation of many contacts unfolding events at a value of d_u indicates emergence of a force peak. In such figure the y-axis shows the sequential separation between two amino acids that make a native contact. The x-axis indicates at which displacement of the pulling tip the contact is broken for the last time. Notice that thermal fluctuations may reinstate a broken contact temporarily and that’s why we seek the last, and thus definitive, rupture event.

As an example we show the unfolding diagram for the first chain of protein 1c4p. This protein contains an α helix (196-210), denoted as I, and eight β strands, denoted as A through H, as labelled consecutively from the N to C termini. These β strands form three β sheets: a, b, and c. The a sheet comprises four strands (A, B, G, E), b sheet two ( C, F), and c-sheet also two ( D, H). These letters are indicated on the unfolding scenario figure and show which secondary structures are involved in a contact that is broken at the distance d_u. The data symbols marked by asterisks correspond to contacts which do not involve any secondary structures. The remaining symbols are diversified and have a meaning identified by the letter symbols placed next to them. The force peak at about 140 Å involves a near simultaneous rupture of the contacts which relate to the terminal strands A and G: A+G (meaning between A and G), A+I, G+I (i.e. with the helix) as well as D+H, A+B, E+G, D+G, C+G, and A+C. Breaking these contacts results in destroying the a β-sheet. Later on, the remaining contacts, such as C+F in the b sheet are broken and the helix is the last to unravel.

ICN I J |I-J| Displacement error 1 697 702 5 809.46 1.35 2 697 703 6 547.78 70.49 3 698 703 5 754.66 1.44 4 699 703 4 821.62 3.26 5 700 703 3 858.76 1.85 6 701 815 114 91.36 1.03 7 701 816 115 87.03 0.70 8 701 817 116 87.03 0.70 9 702 705 3 829.39 0.22 10 702 817 115 94.61 0.71 11 704 707 3 861.46 1.86 12 704 708 4 850.58 1.16 13 704 716 12 99.33 0.75 14 705 708 3 874.59 0.48 15 705 709 4 856.11 1.61 16 705 716 11 99.51 0.81 17 705 798 93 99.32 0.77 18 705 817 112 99.33 0.77 19 705 818 113 99.32 0.77 20 705 819 114 99.20 0.82 21 705 821 116 98.60 0.52 22 706 709 3 871.25 1.19 23 706 937 231 57.94 0.40 24 707 711 4 844.95 5.20 25 708 711 3 846.29 4.53 26 708 713 5 803.50 1.62 27 708 714 6 751.55 0.53 28 708 715 7 744.82 2.84 29 708 716 8 444.60 19.68 30 708 726 18 103.31 0.86 31 708 819 111 99.39 0.81 32 708 824 116 99.27 0.77 33 709 821 112 99.39 0.76 34 709 823 114 99.28 0.83 35 709 824 115 99.25 0.78 36 709 872 163 64.95 0.68 37 709 933 224 68.15 0.30 38 709 937 228 63.51 1.40 39 710 868 158 63.51 1.40 40 710 871 161 63.51 1.40 41 710 872 162 64.95 0.69 42 710 937 227 66.29 0.16 43 711 824 113 99.56 0.73 44 712 726 14 104.11 1.15 45 712 727 15 103.37 0.83 46 712 728 16 99.95 0.83 47 712 729 17 99.89 0.86 48 712 824 112 99.78 0.84 49 713 726 13 104.09 1.16 50 713 727 14 103.36 0.81 51 714 725 11 114.79 2.44 52 714 726 12 114.33 2.33 53 715 718 3 935.87 5.01 54 715 724 9 115.11 2.37 55 715 725 10 114.81 2.44 56 716 724 8 115.17 2.35 57 716 726 10 114.98 2.37 58 716 816 100 115.00 2.41 59 716 817 101 114.98 2.40 60 716 818 102 114.75 2.46 61 716 819 103 113.81 2.93 62 717 723 6 234.22 61.54 63 717 724 7 115.15 2.39 64 717 814 97 256.47 72.67 65 717 816 99 214.07 51.85 66 718 722 4 749.44 0.04 67 718 723 5 223.59 56.22 68 718 744 26 514.36 99.31 69 719 722 3 848.75 4.29 70 719 744 25 514.48 99.32 71 719 814 95 385.50 71.22 72 720 744 24 514.48 99.32 73 721 743 22 514.48 99.32 74 721 744 23 514.48 99.32 75 721 745 24 514.46 99.33 76 721 749 28 512.34 100.31 77 721 805 84 438.36 65.23 78 721 807 86 438.60 65.22 79 721 812 91 437.86 65.14 80 722 741 19 514.52 99.31 81 722 742 20 514.50 99.31 82 722 744 22 514.50 99.31 83 722 749 27 514.38 99.31 84 722 752 30 511.28 100.72 85 722 803 81 437.46 65.09 86 722 805 83 439.12 65.29 87 723 740 17 514.71 99.33 88 723 741 18 514.59 99.29 89 723 742 19 514.57 99.28 90 723 744 21 514.57 99.28 91 724 739 15 514.91 99.27 92 724 740 16 514.71 99.33 93 724 741 17 514.61 99.28 94 724 801 77 442.42 67.62 95 724 803 79 442.59 67.23 96 724 816 92 437.62 65.22 97 725 739 14 515.01 99.22 98 725 740 15 514.74 99.31 99 726 737 11 515.23 99.29 100 726 738 12 515.03 99.23 101 726 739 13 515.02 99.22 102 726 740 14 515.01 99.22 103 726 818 92 437.46 65.12 104 726 819 93 433.73 65.94 105 727 737 10 515.34 99.24 106 727 738 11 515.14 99.25 107 727 740 13 515.08 99.22 108 728 736 8 515.35 99.24 109 728 737 9 515.33 99.24 110 729 735 6 515.48 99.24 111 729 736 7 515.41 99.22 112 730 734 4 721.30 3.66 113 730 735 5 515.48 99.23 114 730 789 59 515.60 98.67 115 730 824 94 416.38 59.75 116 730 825 95 415.64 59.77 117 731 734 3 835.65 3.12 118 732 825 93 416.61 59.81 119 732 894 162 297.22 0.48 120 733 790 57 557.04 77.94 121 733 791 58 556.71 77.92 122 733 825 92 416.61 59.81 123 733 894 161 297.55 0.33 124 733 895 162 301.22 0.97 125 733 896 163 303.28 1.45 126 734 789 55 557.46 77.75 127 734 790 56 556.90 77.80 128 734 825 91 416.59 59.83 129 735 788 53 557.88 77.55 130 735 789 54 557.62 77.44 131 736 786 50 558.18 77.45 132 736 787 51 557.97 77.50 133 736 788 52 557.87 77.54 134 736 789 53 557.64 77.43 135 737 786 49 558.15 77.41 136 737 787 50 557.96 77.48 137 737 789 52 557.73 77.39 138 737 818 81 438.46 65.22 139 737 819 82 438.20 65.17 140 737 820 83 433.39 65.34 141 738 784 46 558.41 77.34 142 738 785 47 558.20 77.38 143 738 786 48 558.17 77.38 144 739 784 45 558.38 77.31 145 739 785 46 558.29 77.33 146 739 787 48 558.17 77.36 147 739 801 62 489.82 44.01 148 739 818 79 437.67 65.22 149 740 782 42 558.68 77.20 150 740 783 43 558.54 77.23 151 740 784 44 558.37 77.29 152 741 752 11 558.59 77.20 153 741 754 13 558.10 77.13 154 741 782 41 558.66 77.19 155 741 783 42 558.58 77.20 156 741 785 44 558.50 77.20 157 741 801 60 490.18 43.74 158 741 803 62 489.61 43.72 159 742 781 39 559.71 76.69 160 742 782 40 558.60 77.15 161 743 747 4 841.81 1.96 162 743 748 5 746.27 0.64 163 743 749 6 696.94 8.18 164 743 777 34 562.84 75.21 165 743 778 35 562.88 75.20 166 743 779 36 562.87 75.20 167 743 780 37 562.86 75.20 168 743 781 38 560.61 76.17 169 745 779 34 593.37 68.92 170 745 780 35 590.75 69.90 171 745 807 62 489.00 43.77 172 746 779 33 601.75 69.69 173 747 779 32 604.58 68.38 174 747 807 60 488.96 43.80 175 748 776 28 606.43 67.67 176 748 777 29 604.76 68.45 177 748 778 30 604.75 68.45 178 748 779 31 605.91 67.71 179 749 752 3 880.95 0.84 180 749 776 27 606.48 67.69 181 749 777 28 606.30 67.72 182 749 805 56 489.73 43.72 183 749 806 57 489.29 43.66 184 749 807 58 488.97 43.80 185 750 775 25 606.83 67.84 186 750 776 26 606.50 67.69 187 750 805 55 489.79 43.74 188 750 806 56 489.34 43.90 189 751 775 24 607.50 68.17 190 751 804 53 490.00 43.73 191 751 805 54 489.45 43.91 192 751 806 55 489.43 43.92 193 752 773 21 607.49 68.17 194 752 775 23 607.51 68.19 195 752 777 25 607.61 68.21 196 752 783 31 607.63 68.20 197 752 803 51 490.06 43.72 198 752 804 52 489.88 43.78 199 752 805 53 489.41 43.93 200 753 803 50 490.09 43.71 201 753 804 51 489.89 43.79 202 754 773 19 607.66 68.18 203 754 785 31 607.62 68.20 204 754 801 47 490.54 43.79 205 754 802 48 490.42 43.75 206 754 803 49 490.11 43.70 207 755 801 46 490.54 43.79 208 755 802 47 490.38 43.73 209 756 768 12 607.44 68.28 210 756 770 14 607.72 68.23 211 756 787 31 607.71 68.23 212 756 800 44 490.65 43.83 213 756 801 45 490.55 43.79 214 756 818 62 446.00 64.84 215 757 799 42 490.75 43.87 216 757 800 43 490.66 43.83 217 757 802 45 490.62 43.83 218 758 762 4 829.58 0.48 219 758 763 5 817.36 3.11 220 758 768 10 608.94 68.11 221 758 799 41 490.76 43.87 222 759 762 3 855.18 5.69 223 759 763 4 834.82 2.04 224 759 797 38 491.40 43.90 225 760 763 3 855.70 5.44 226 760 764 4 844.44 0.20 227 761 764 3 851.96 3.41 228 761 765 4 845.47 0.17 229 761 797 36 537.35 46.40 230 762 765 3 845.47 0.17 231 762 766 4 840.77 2.03 232 762 795 33 581.01 66.87 233 762 797 35 511.94 47.31 234 762 798 36 491.92 43.59 235 762 799 37 491.16 43.96 236 763 766 3 853.10 3.98 237 764 792 28 607.72 68.24 238 765 791 26 607.73 68.25 239 765 792 27 607.70 68.25 240 765 793 28 607.61 68.22 241 765 794 29 607.60 68.22 242 765 795 30 602.39 66.08 243 766 789 23 607.91 68.18 244 766 790 24 607.80 68.22 245 766 791 25 607.72 68.25 246 766 792 26 607.72 68.25 247 766 795 29 596.08 71.34 248 766 799 33 490.73 43.89 249 766 820 54 440.39 64.66 250 766 825 59 429.67 64.64 251 767 789 22 607.95 68.17 252 767 790 23 607.89 68.17 253 767 792 25 607.74 68.25 254 768 787 19 608.28 68.12 255 768 788 20 608.18 68.10 256 768 789 21 607.99 68.19 257 768 799 31 490.48 43.99 258 768 820 52 438.83 65.44 259 769 787 18 608.28 68.12 260 769 788 19 608.25 68.12 261 770 785 15 608.41 68.14 262 770 786 16 608.38 68.13 263 770 787 17 608.28 68.12 264 771 786 15 608.40 68.14 265 773 783 10 608.63 68.16 266 773 784 11 608.60 68.15 267 773 785 12 608.43 68.14 268 774 783 9 608.64 68.17 269 777 781 4 777.43 1.51 270 777 782 5 734.78 5.36 271 777 783 6 718.69 13.28 272 778 781 3 804.02 11.79 273 785 801 16 490.23 43.90 274 789 820 31 438.23 65.16 275 789 825 36 417.41 59.41 276 790 896 106 303.42 1.51 277 791 795 4 854.52 1.50 278 791 825 34 421.63 60.66 279 791 895 104 302.45 1.17 280 791 896 105 303.42 1.51 281 792 895 103 303.36 1.48 282 792 896 104 303.43 1.51 283 792 897 105 303.44 1.51 284 793 895 102 303.53 1.49 285 793 897 104 303.56 1.47 286 794 822 28 438.86 65.42 287 794 895 101 307.08 0.28 288 794 923 129 187.29 0.28 289 794 925 131 160.14 0.45 290 794 926 132 160.08 0.46 291 795 820 25 445.55 64.87 292 795 821 26 441.92 65.36 293 795 822 27 438.42 65.20 294 795 825 30 438.14 65.20 295 795 929 134 158.44 0.19 296 796 929 133 156.56 0.51 297 798 818 20 446.44 65.06 298 799 817 18 446.45 65.06 299 799 818 19 446.37 65.07 300 799 820 21 444.74 64.46 301 800 815 15 446.62 65.00 302 800 816 16 446.49 65.05 303 800 817 17 446.43 65.05 304 801 815 14 446.62 64.98 305 801 816 15 446.58 64.99 306 801 818 17 445.01 64.34 307 802 813 11 446.85 64.98 308 802 814 12 446.67 64.98 309 802 815 13 446.58 64.98 310 803 813 10 446.76 64.94 311 803 814 11 446.64 64.97 312 804 811 7 447.12 64.86 313 804 812 8 446.86 64.90 314 804 813 9 446.74 64.92 315 805 811 6 447.07 64.84 316 805 812 7 446.91 64.92 317 806 810 4 732.44 10.87 318 807 810 3 839.26 3.02 319 807 812 5 642.75 32.98 320 821 824 3 867.47 4.24 321 821 825 4 859.82 3.56 322 821 929 108 156.65 0.50 323 821 933 112 156.61 0.51 324 822 825 3 876.00 0.99 325 822 893 71 453.51 73.84 326 822 926 104 158.85 0.03 327 822 929 107 157.72 0.07 328 822 930 108 156.65 0.51 329 822 933 111 156.64 0.50 330 823 826 3 865.15 0.90 331 823 872 49 411.95 60.91 332 823 893 70 459.49 76.67 333 823 930 107 157.72 0.07 334 823 933 110 157.71 0.07 335 825 892 67 469.50 81.07 336 825 893 68 469.38 81.02 337 825 894 69 469.40 81.02 338 826 872 46 468.98 81.20 339 826 873 47 467.88 80.86 340 826 874 48 468.34 80.62 341 826 891 65 469.34 81.03 342 826 892 66 469.48 81.06 343 826 893 67 469.48 81.06 344 826 902 76 469.41 81.03 345 826 930 104 171.66 0.57 346 827 891 64 469.51 81.07 347 827 892 65 469.51 81.07 348 827 894 67 469.62 81.09 349 828 873 45 469.51 81.09 350 828 874 46 469.48 81.08 351 828 875 47 469.47 81.08 352 828 890 62 469.53 81.06 353 828 891 63 469.54 81.06 354 829 875 46 469.57 81.06 355 829 877 48 469.61 81.06 356 829 890 61 469.57 81.06 357 829 891 62 469.57 81.06 358 829 892 63 469.63 81.08 359 830 875 45 469.62 81.08 360 830 877 47 469.62 81.07 361 831 835 4 837.49 0.97 362 831 836 5 829.98 2.71 363 831 839 8 798.26 0.97 364 831 877 46 469.64 81.06 365 831 890 59 469.69 81.06 366 832 835 3 846.51 3.53 367 832 836 4 846.51 3.54 368 833 836 3 848.12 4.25 369 833 837 4 846.52 3.52 370 833 880 47 469.68 81.08 371 834 837 3 850.50 3.06 372 834 838 4 837.51 0.98 373 835 838 3 848.99 2.30 374 835 839 4 839.51 0.02 375 836 839 3 854.08 2.00 376 836 840 4 837.37 1.05 377 836 877 41 470.71 81.34 378 836 880 44 470.73 81.33 379 836 888 52 470.57 81.31 380 836 889 53 470.62 81.29 381 836 903 67 470.72 81.36 382 837 840 3 853.73 2.17 383 837 841 4 835.58 0.10 384 837 880 43 470.80 81.34 385 837 881 44 470.81 81.34 386 837 882 45 470.85 81.34 387 838 841 3 853.74 2.17 388 838 842 4 843.00 3.21 389 839 842 3 852.44 5.71 390 839 903 64 470.80 81.33 391 840 843 3 887.98 1.19 392 840 903 63 470.81 81.33 393 840 904 64 470.80 81.33 394 840 905 65 470.81 81.33 395 840 918 78 373.32 71.70 396 840 920 80 357.42 69.00 397 841 920 79 369.56 73.48 398 843 846 3 940.89 1.55 399 843 848 5 772.81 11.89 400 843 849 6 745.10 1.96 401 843 920 77 369.66 73.50 402 845 848 3 864.72 1.61 403 846 849 3 859.49 0.95 404 847 924 77 368.45 73.69 405 848 919 71 424.68 81.75 406 848 920 72 409.29 81.36 407 848 921 73 405.12 82.58 408 848 922 74 387.34 73.70 409 848 923 75 369.20 73.34 410 848 924 76 353.29 70.56 411 848 927 79 198.97 2.15 412 849 918 69 430.62 80.57 413 849 919 70 424.68 81.75 414 849 920 71 409.55 81.23 415 850 918 68 430.64 80.56 416 850 919 69 426.37 82.59 417 850 927 77 192.89 0.35 418 850 928 78 192.80 0.39 419 850 931 81 192.41 0.24 420 851 916 65 445.99 86.68 421 851 917 66 445.53 86.88 422 851 918 67 434.76 82.62 423 852 916 64 460.32 81.13 424 852 917 65 445.88 86.73 425 852 931 79 192.57 0.24 426 852 935 83 190.81 0.55 427 853 914 61 465.69 80.31 428 853 915 62 462.24 82.00 429 853 916 63 458.72 80.33 430 854 914 60 465.71 80.31 431 854 915 61 462.30 82.00 432 854 917 63 462.24 82.00 433 854 935 81 191.27 0.54 434 854 938 84 189.78 0.63 435 855 913 58 466.59 79.88 436 855 914 59 462.29 82.01 437 856 860 4 815.68 3.93 438 856 913 57 465.71 80.30 439 856 914 58 462.29 82.00 440 856 915 59 462.29 82.01 441 856 938 82 189.38 0.45 442 857 938 81 189.38 0.45 443 858 868 10 559.85 68.89 444 858 937 79 189.75 0.62 445 858 938 80 189.20 0.43 446 860 908 48 471.84 81.14 447 860 909 49 471.85 81.16 448 860 910 50 471.69 81.17 449 860 913 53 447.62 86.95 450 860 914 54 447.62 86.95 451 860 915 55 442.88 86.25 452 861 864 3 841.10 2.62 453 861 865 4 837.21 4.04 454 862 865 3 839.71 2.80 455 862 866 4 839.33 2.99 456 862 887 25 494.69 70.06 457 863 866 3 839.70 2.79 458 863 867 4 833.94 0.27 459 864 867 3 841.04 2.51 460 864 868 4 834.32 0.07 461 865 868 3 858.07 3.35 462 865 869 4 834.33 0.07 463 865 887 22 495.12 69.85 464 865 906 41 472.35 81.18 465 865 938 73 183.75 0.80 466 866 869 3 863.25 0.76 467 866 870 4 845.87 0.10 468 866 876 10 510.38 67.64 469 866 878 12 502.65 73.59 470 866 887 21 495.12 69.84 471 867 870 3 856.16 5.24 472 867 871 4 836.31 4.69 473 868 871 3 860.00 5.17 474 868 872 4 836.47 4.61 475 868 937 69 184.28 0.55 476 868 938 70 183.76 0.80 477 869 872 3 848.18 1.15 478 869 873 4 822.40 2.79 479 869 874 5 816.39 0.96 480 869 876 7 550.50 67.60 481 869 887 18 495.22 69.81 482 869 906 37 472.50 81.17 483 869 938 69 183.60 0.72 484 870 873 3 858.00 6.17 485 870 876 6 559.17 63.27 486 872 930 58 190.75 0.50 487 872 933 61 189.22 0.53 488 872 934 62 187.50 1.28 489 874 889 15 505.97 75.19 490 874 890 16 505.95 75.20 491 874 891 17 505.88 75.23 492 874 904 30 483.51 86.42 493 875 889 14 506.14 75.17 494 875 890 15 506.04 75.23 495 876 887 11 506.23 75.13 496 876 888 12 506.23 75.13 497 876 889 13 506.22 75.14 498 877 880 3 874.23 3.68 499 877 888 11 506.23 75.13 500 877 890 13 506.31 75.18 501 880 888 8 506.55 75.20 502 881 886 5 528.46 65.06 503 881 888 7 506.60 75.22 504 881 905 24 483.32 86.88 505 883 886 3 865.27 2.69 506 885 908 23 483.20 86.95 507 886 905 19 483.57 86.99 508 886 906 20 483.36 86.92 509 886 907 21 483.17 86.97 510 886 908 22 483.20 86.95 511 887 905 18 483.57 86.98 512 887 906 19 483.47 87.00 513 888 904 16 483.88 86.87 514 888 905 17 483.57 86.98 515 889 903 14 484.38 86.89 516 889 904 15 484.11 86.97 517 890 902 12 485.62 86.50 518 890 903 13 484.62 86.73 519 891 901 10 486.72 86.56 520 891 902 11 486.60 86.53 521 892 899 7 514.57 72.84 522 892 900 8 508.81 75.70 523 892 901 9 486.72 86.56 524 893 899 6 515.90 73.47 525 893 900 7 514.56 72.85 526 893 902 9 487.18 86.48 527 893 926 33 189.16 1.15 528 893 930 37 186.85 2.05 529 894 899 5 515.91 73.47 530 895 898 3 842.50 0.58 531 895 899 4 640.12 76.30 532 900 921 21 192.82 0.39 533 900 922 22 192.71 0.37 534 900 923 23 192.66 0.36 535 900 926 26 192.43 0.32 536 901 921 20 192.83 0.38 537 901 922 21 192.75 0.40 538 902 920 18 192.93 0.41 539 902 921 19 192.81 0.39 540 902 926 24 192.45 0.30 541 902 930 28 191.15 0.48 542 903 919 16 195.99 0.54 543 903 920 17 193.47 0.64 544 904 917 13 367.53 73.80 545 904 918 14 210.08 2.71 546 904 919 15 195.97 0.55 547 904 934 30 189.55 0.53 548 905 917 12 367.60 73.84 549 905 918 13 362.45 71.42 550 906 915 9 382.84 76.45 551 906 916 10 369.68 73.46 552 906 917 11 367.66 73.86 553 906 938 32 183.46 0.65 554 907 915 8 383.48 76.14 555 907 916 9 369.76 73.48 556 908 914 6 516.10 73.99 557 908 915 7 394.06 81.42 558 909 912 3 841.78 4.51 559 909 913 4 640.84 79.86 560 909 914 5 542.50 76.38 561 910 913 3 807.99 3.37 562 915 938 23 189.11 0.48 563 917 931 14 192.13 0.18 564 917 934 17 191.04 0.66 565 917 935 18 189.56 0.54 566 919 927 8 486.66 87.47 567 919 930 11 192.40 0.25 568 919 931 12 191.33 0.57 569 921 926 5 782.54 6.16 570 921 927 6 558.08 69.17 571 921 930 9 560.07 70.17 572 923 926 3 848.81 3.36 573 923 927 4 831.31 5.39 574 924 927 3 855.79 2.68 575 924 928 4 831.83 5.65 576 925 928 3 848.11 6.52 577 925 929 4 831.86 5.65 578 926 929 3 848.92 5.82 579 926 930 4 831.88 5.63 580 927 930 3 861.69 0.98 581 927 931 4 831.88 5.63 582 928 931 3 855.18 4.24 583 928 932 4 837.86 2.65 584 929 932 3 846.90 3.20 585 929 933 4 831.28 4.44 586 930 933 3 842.01 0.15 587 930 934 4 843.47 1.54 588 931 934 3 845.70 0.43 589 931 935 4 843.59 1.49 590 932 935 3 855.25 2.14 591 932 936 4 843.87 1.47 592 933 936 3 868.48 2.15 593 933 937 4 857.59 3.30 594 934 937 3 873.63 0.43 595 934 938 4 865.25 0.53 596 935 938 3 874.23 0.14