0 11 Involvement involvement NN 12 14 of of IN 15 18 the the DT 19 29 N-terminal n-terminal JJ 30 36 region region NN 37 39 of of IN 40 43 the the DT 44 49 human human JJ 50 67 mineralocorticoid mineralocorticoid NN 68 76 receptor receptor NN 77 92 hormone-binding hormone-binding JJ 93 99 domain domain NN 100 102 in in IN 103 110 agonist agonist NN 111 114 and and CC 115 125 antagonist antagonist NN 126 133 binding binding NN 134 136 as as IN 137 145 revealed reveal VBN 146 148 by by IN 149 150 a a DT 151 154 new new JJ 155 165 monoclonal monoclonal JJ 166 174 antibody antibody NN 174 175 . . . 177 179 To to TO 180 184 gain gain VB 185 186 a a DT 187 193 better good JJR 194 207 understanding understanding NN 208 210 of of IN 211 214 the the DT 215 224 mechanism mechanism NN 225 227 of of IN 228 235 binding bind VBG 236 238 to to TO 239 242 the the DT 243 248 human human JJ 249 266 mineralocorticoid mineralocorticoid NN 267 275 receptor receptor NN 276 277 ( ( ( 277 280 hMR hMR NNP 280 281 ) ) ) 281 282 , , , 283 285 we we PRP 286 295 developed develop VBD 296 297 a a DT 298 301 new new JJ 302 312 monoclonal monoclonal JJ 313 321 antibody antibody NN 322 323 ( ( ( 323 326 mAb mab NN 326 327 ) ) ) 328 334 raised raise VBN 335 342 against against IN 343 346 the the DT 347 362 hormone-binding hormone-binding JJ 363 369 domain domain NN 370 371 ( ( ( 371 374 HBD HBD NNP 374 375 ) ) ) 375 376 . . . 377 380 For for IN 381 385 this this DT 386 393 purpose purpose NN 393 394 , , , 395 399 mice mouse NNS 400 404 were be VBD 405 414 immunized immunize VBN 415 419 with with IN 420 421 a a DT 422 428 fusion fusion NN 429 436 protein protein NN 437 446 including include VBG 447 450 the the DT 451 459 sequence sequence NN 460 473 Thr729-Lys984 thr729-lys984 NN 474 476 of of IN 477 480 hMR hMR NNP 480 481 . . . 482 487 After after IN 488 493 ELISA ELISA NNP 494 503 screening screening NN 503 504 , , , 505 508 mAb mab NN 509 513 18C7 18c7 NN 514 517 was be VBD 518 526 selected select VBN 527 530 for for IN 531 534 its its PRP$ 535 546 specificity specificity NN 547 554 towards towards IN 555 558 the the DT 559 562 HBD HBD NNP 562 563 . . . 564 568 This this DT 569 577 antibody antibody NN 578 588 recognized recognize VBD 589 593 both both CC 594 597 the the DT 598 607 denatured denature VBN 608 611 and and CC 612 618 native native JJ 619 621 MR mr NN 622 627 forms form NNS 627 628 , , , 629 631 as as RB 632 636 well well RB 637 639 as as IN 640 643 the the DT 644 661 hetero-oligomeric hetero-oligomeric JJ 662 664 MR mr NN 665 669 form form NN 670 673 and and CC 674 677 the the DT 678 689 transformed transform VBN 690 692 MR mr NN 693 698 state state NN 698 699 . . . 700 702 By by IN 703 708 using use VBG 709 716 several several JJ 717 720 HBD HBD NNP 721 733 subfragments subfragment NNS 733 734 , , , 735 738 the the DT 739 742 mAb mab NN 743 747 18C7 18c7 NN 748 755 epitope epitope NN 756 759 was be VBD 760 767 located locate VBN 768 770 in in IN 771 774 the the DT 775 785 N-terminal n-terminal JJ 786 792 region region NN 793 795 of of IN 796 799 the the DT 800 803 HBD hbd NN 804 808 from from IN 809 815 Thr729 thr729 NN 816 818 to to TO 819 825 Leu765 leu765 NN 825 826 . . . 827 829 We we PRP 830 834 then then RB 835 842 studied study VBD 843 846 the the DT 847 853 effect effect NN 854 856 of of IN 857 860 the the DT 861 869 antibody antibody NN 870 872 on on IN 873 884 aldosterone aldosterone NN 885 888 and and CC 889 901 progesterone progesterone NN 902 909 binding binding NN 910 912 to to TO 913 916 the the DT 917 920 hMR hMR NNP 920 921 . . . 922 926 When when WRB 927 931 18C7 18c7 NN 932 935 was be VBD 936 945 incubated incubate VBN 946 950 with with IN 951 959 liganded liganded JJ 960 962 MR mr NN 962 963 , , , 964 966 it it PRP 967 970 was be VBD 971 975 able able JJ 976 978 to to TO 979 985 partly partly RB 986 994 displace displace VB 995 996 ( ( ( 996 998 20 20 CD 998 999 % % NN 999 1000 ) ) ) 1001 1004 the the DT 1005 1012 hormone hormone NN 1013 1017 from from IN 1018 1021 its its PRP$ 1022 1029 binding bind VBG 1030 1034 site site NN 1034 1035 . . . 1036 1040 When when WRB 1041 1045 18C7 18c7 NN 1046 1049 was be VBD 1050 1059 incubated incubate VBN 1060 1064 with with IN 1065 1067 MR mr NN 1068 1074 before before IN 1075 1086 aldosterone aldosterone NN 1087 1089 or or CC 1090 1102 progesterone progesterone NN 1102 1103 , , , 1104 1107 the the DT 1108 1116 antibody antibody NN 1117 1126 inhibited inhibit VBD 1127 1132 75-80 75-80 CD 1132 1133 % % NN 1134 1136 of of IN 1137 1140 the the DT 1141 1148 binding binding NN 1148 1149 . . . 1150 1153 The the DT 1154 1160 effect effect NN 1161 1163 of of IN 1164 1168 18C7 18c7 NN 1169 1171 on on IN 1172 1175 the the DT 1176 1183 binding binding NN 1184 1187 was be VBD 1188 1195 similar similar JJ 1196 1200 with with IN 1201 1205 both both DT 1206 1214 hormones hormone NNS 1214 1215 . . . 1216 1217 A a DT 1218 1225 sucrose sucrose NN 1226 1234 gradient gradient NN 1235 1243 analysis analysis NN 1244 1253 indicated indicate VBD 1254 1257 the the DT 1258 1270 simultaneous simultaneous JJ 1271 1279 presence presence NN 1280 1282 of of IN 1283 1286 two two CD 1287 1292 kinds kind NNS 1293 1295 of of IN 1296 1304 receptor receptor NN 1305 1314 complexes complex NNS 1314 1315 : : : 1316 1319 the the DT 1320 1330 steroid-MR steroid-MR NNP 1331 1338 complex complex NN 1339 1342 and and CC 1343 1346 the the DT 1347 1358 antibody-MR antibody-mr JJ 1359 1366 complex complex NN 1366 1367 . . . 1368 1373 After after IN 1374 1377 its its PRP$ 1378 1388 associated associate VBN 1389 1397 proteins protein NNS 1397 1398 , , , 1399 1409 especially especially RB 1410 1413 the the DT 1414 1424 heat-shock heat-shock NN 1425 1432 protein protein NN 1433 1438 hsp90 hsp90 NN 1438 1439 , , , 1440 1443 had have VBD 1444 1448 been be VBN 1449 1461 cross-linked cross-linked JJ 1462 1466 with with IN 1467 1470 the the DT 1471 1474 hMR hMR NNP 1475 1477 by by IN 1478 1498 dimethylpimelimidate dimethylpimelimidate NN 1498 1499 , , , 1500 1504 18C7 18c7 NN 1505 1508 was be VBD 1509 1514 still still RB 1515 1519 able able JJ 1520 1522 to to TO 1523 1528 react react VB 1529 1533 with with IN 1534 1537 the the DT 1538 1546 receptor receptor NN 1546 1547 . . . 1548 1551 Our our PRP$ 1552 1559 results result NNS 1560 1569 indicated indicate VBD 1570 1574 that that IN 1575 1578 the the DT 1579 1586 epitope epitope NN 1587 1597 recognized recognize VBN 1598 1600 by by IN 1601 1605 18C7 18c7 NN 1606 1609 was be VBD 1610 1618 directly directly RB 1619 1629 implicated implicate VBN 1630 1632 in in IN 1633 1640 hormone hormone NN 1641 1648 binding binding NN 1648 1649 . . . 1650 1653 The the DT 1654 1658 lack lack NN 1659 1661 of of IN 1662 1669 steroid steroid NN 1670 1677 binding binding NN 1678 1680 of of IN 1681 1684 HBD hbd NN 1685 1692 mutants mutant NNS 1693 1697 with with IN 1698 1701 the the DT 1702 1715 Thr729-Leu765 thr729-leu765 JJ 1716 1724 sequence sequence NN 1725 1732 deleted delete VBN 1733 1734 [ [ ( 1734 1743 Jalaguier Jalaguier NNP 1743 1744 , , , 1745 1752 Mesnier Mesnier NNP 1752 1753 , , , 1754 1759 Leger Leger NNP 1760 1763 and and CC 1764 1769 Auzou Auzou NNP 1770 1771 ( ( ( 1771 1775 1996 1996 CD 1775 1776 ) ) ) 1777 1786 J.Steroid J.Steroid NNP 1787 1806 Biochem.Mol.Biol.57 Biochem.Mol.Biol.57 NNP 1806 1807 , , , 1808 1813 43-50 43-50 CD 1813 1814 ] ] ) 1815 1823 supports support VBZ 1824 1828 this this DT 1829 1839 hypothesis hypothesis NN 1839 1840 . . . 1841 1848 Because because IN 1849 1851 of of IN 1852 1855 the the DT 1856 1863 similar similar JJ 1864 1874 behaviours behaviour NNS 1875 1877 of of IN 1878 1889 aldosterone aldosterone NN 1890 1893 and and CC 1894 1906 progesterone progesterone NN 1906 1907 , , , 1908 1910 we we PRP 1911 1919 conclude conclude VBP 1920 1924 that that IN 1925 1928 the the DT 1929 1939 N-terminal n-terminal JJ 1940 1953 Thr729-Leu765 Thr729-Leu765 NNP 1954 1960 region region NN 1961 1963 of of IN 1964 1967 the the DT 1968 1971 HBD HBD NNP 1972 1974 is be VBZ 1975 1984 similarly similarly RB 1985 1993 involved involve VBN 1994 1996 in in IN 1997 2000 the the DT 2001 2008 binding binding NN 2009 2011 of of IN 2012 2016 both both DT 2017 2025 hormones hormone NNS 2025 2026 . . .