HEADER IMMUNE SYSTEM 10-DEC-24 9MFN TITLE CRYSTAL STRUCTURE OF ADI-64596 (HUMAN FAB, WITH SUBSTITUTED IGG1-CH1 TITLE 2 (HC-L145Q, K147E, AND S181E) AND SUBSTITUTED KAPPA CONSTANT DOMAIN TITLE 3 (LC-T129R, T178R, AND T180Q)) COMPND MOL_ID: 1; COMPND 2 MOLECULE: ADI-64596 FAB HEAVY CHAIN; COMPND 3 CHAIN: H; COMPND 4 ENGINEERED: YES; COMPND 5 OTHER_DETAILS: SUBSTITUTED IGG1-CH1 (HC-L145Q, K147E, AND S181E; COMPND 6 MOL_ID: 2; COMPND 7 MOLECULE: ADI-64596 FAB LIGHT CHAIN; COMPND 8 CHAIN: L; COMPND 9 ENGINEERED: YES; COMPND 10 OTHER_DETAILS: SUBSTITUTED KAPPA CONSTANT DOMAIN (LC-T129R, T178R, COMPND 11 AND T180Q SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 3 ORGANISM_COMMON: HUMAN; SOURCE 4 ORGANISM_TAXID: 9606; SOURCE 5 EXPRESSION_SYSTEM: CRICETULUS GRISEUS; SOURCE 6 EXPRESSION_SYSTEM_COMMON: CHINESE HAMSTER; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 10029; SOURCE 8 MOL_ID: 2; SOURCE 9 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 10 ORGANISM_COMMON: HUMAN; SOURCE 11 ORGANISM_TAXID: 9606; SOURCE 12 EXPRESSION_SYSTEM: CRICETULUS GRISEUS; SOURCE 13 EXPRESSION_SYSTEM_COMMON: CHINESE HAMSTER; SOURCE 14 EXPRESSION_SYSTEM_TAXID: 10029 KEYWDS ANTIBODY, IMMUNE SYSTEM EXPDTA X-RAY DIFFRACTION AUTHOR M.B.BATTLES,M.WELIN,M.LAURSEN REVDAT 1 02-APR-25 9MFN 0 JRNL AUTH K.A.BARLOW,M.B.BATTLES,M.E.BROWN,K.CANFIELD,X.LU,H.LYNAUGH, JRNL AUTH 2 M.MORRILL,C.G.RAPPAZZO,S.P.REYES,C.SANDBERG,B.SHARKEY, JRNL AUTH 3 C.STRONG,J.ZHAO,A.SIVASUBRAMANIAN JRNL TITL DESIGN OF ORTHOGONAL CONSTANT DOMAIN INTERFACES TO AID JRNL TITL 2 PROPER HEAVY/LIGHT CHAIN PAIRING OF BISPECIFIC ANTIBODIES. JRNL REF MABS V. 17 79531 2025 JRNL REFN ESSN 1942-0870 JRNL PMID 40126074 JRNL DOI 10.1080/19420862.2025.2479531 REMARK 2 REMARK 2 RESOLUTION. 2.35 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : PHENIX (1.18.2_3874: ???) REMARK 3 AUTHORS : PAUL ADAMS,PAVEL AFONINE,VINCENT CHEN,IAN REMARK 3 : DAVIS,KRESHNA GOPAL,RALF GROSSE-KUNSTLEVE, REMARK 3 : LI-WEI HUNG,ROBERT IMMORMINO,TOM IOERGER, REMARK 3 : AIRLIE MCCOY,ERIK MCKEE,NIGEL MORIARTY, REMARK 3 : REETAL PAI,RANDY READ,JANE RICHARDSON, REMARK 3 : DAVID RICHARDSON,TOD ROMO,JIM SACCHETTINI, REMARK 3 : NICHOLAS SAUTER,JACOB SMITH,LAURENT REMARK 3 : STORONI,TOM TERWILLIGER,PETER ZWART REMARK 3 REMARK 3 REFINEMENT TARGET : ML REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 2.35 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 48.99 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.960 REMARK 3 COMPLETENESS FOR RANGE (%) : 100.0 REMARK 3 NUMBER OF REFLECTIONS : 19398 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.209 REMARK 3 R VALUE (WORKING SET) : 0.208 REMARK 3 FREE R VALUE : 0.221 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.530 REMARK 3 FREE R VALUE TEST SET COUNT : 879 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT (IN BINS). REMARK 3 BIN RESOLUTION RANGE COMPL. NWORK NFREE RWORK RFREE REMARK 3 1 48.9900 - 4.2700 1.00 3049 196 0.1472 0.1528 REMARK 3 2 4.2600 - 3.3900 1.00 3065 156 0.1909 0.2212 REMARK 3 3 3.3900 - 2.9600 1.00 3093 141 0.2655 0.2978 REMARK 3 4 2.9600 - 2.6900 1.00 3146 108 0.2631 0.4012 REMARK 3 5 2.6900 - 2.5000 1.00 3076 144 0.3097 0.3305 REMARK 3 6 2.5000 - 2.3500 1.00 3090 134 0.3317 0.3764 REMARK 3 REMARK 3 BULK SOLVENT MODELLING. REMARK 3 METHOD USED : FLAT BULK SOLVENT MODEL REMARK 3 SOLVENT RADIUS : 1.11 REMARK 3 SHRINKAGE RADIUS : 0.90 REMARK 3 K_SOL : NULL REMARK 3 B_SOL : NULL REMARK 3 REMARK 3 ERROR ESTIMATES. REMARK 3 COORDINATE ERROR (MAXIMUM-LIKELIHOOD BASED) : 0.380 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 32.610 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : NULL REMARK 3 MEAN B VALUE (OVERALL, A**2) : NULL REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : NULL REMARK 3 B22 (A**2) : NULL REMARK 3 B33 (A**2) : NULL REMARK 3 B12 (A**2) : NULL REMARK 3 B13 (A**2) : NULL REMARK 3 B23 (A**2) : NULL REMARK 3 REMARK 3 TWINNING INFORMATION. REMARK 3 FRACTION: NULL REMARK 3 OPERATOR: NULL REMARK 3 REMARK 3 DEVIATIONS FROM IDEAL VALUES. REMARK 3 RMSD COUNT REMARK 3 BOND : 0.002 3347 REMARK 3 ANGLE : 0.551 4558 REMARK 3 CHIRALITY : 0.044 517 REMARK 3 PLANARITY : 0.004 585 REMARK 3 DIHEDRAL : 12.504 1207 REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : 13 REMARK 3 TLS GROUP : 1 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 2 THROUGH 33 ) REMARK 3 ORIGIN FOR THE GROUP (A): 2.6643 -7.2653 -9.4160 REMARK 3 T TENSOR REMARK 3 T11: 0.3918 T22: 0.7217 REMARK 3 T33: 0.7280 T12: 0.0819 REMARK 3 T13: -0.1161 T23: -0.2260 REMARK 3 L TENSOR REMARK 3 L11: 6.0298 L22: 3.5433 REMARK 3 L33: 2.7658 L12: -1.7868 REMARK 3 L13: -3.0480 L23: 2.7239 REMARK 3 S TENSOR REMARK 3 S11: -0.6648 S12: -0.2229 S13: 0.9516 REMARK 3 S21: 0.2017 S22: -0.3652 S23: 0.1453 REMARK 3 S31: -0.0823 S32: -1.2403 S33: 0.8453 REMARK 3 TLS GROUP : 2 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 34 THROUGH 53 ) REMARK 3 ORIGIN FOR THE GROUP (A): 11.2684 -1.5554 -4.6267 REMARK 3 T TENSOR REMARK 3 T11: 0.4854 T22: 0.4876 REMARK 3 T33: 1.0469 T12: 0.0512 REMARK 3 T13: -0.1938 T23: -0.1803 REMARK 3 L TENSOR REMARK 3 L11: 3.7897 L22: 7.9902 REMARK 3 L33: 5.7720 L12: -4.6574 REMARK 3 L13: 3.0868 L23: -1.1438 REMARK 3 S TENSOR REMARK 3 S11: 0.0357 S12: -0.1116 S13: 2.3248 REMARK 3 S21: -0.2036 S22: -1.1822 S23: -0.8011 REMARK 3 S31: -0.9174 S32: -0.3988 S33: 0.3365 REMARK 3 TLS GROUP : 3 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 54 THROUGH 74 ) REMARK 3 ORIGIN FOR THE GROUP (A): 4.7932 5.6356 -7.9006 REMARK 3 T TENSOR REMARK 3 T11: 0.8721 T22: 0.6497 REMARK 3 T33: 1.8348 T12: 0.3846 REMARK 3 T13: -0.5796 T23: -0.3157 REMARK 3 L TENSOR REMARK 3 L11: 1.2396 L22: 0.7596 REMARK 3 L33: 1.7632 L12: -0.4146 REMARK 3 L13: 1.3970 L23: -0.1580 REMARK 3 S TENSOR REMARK 3 S11: -0.7306 S12: -0.4536 S13: 1.1663 REMARK 3 S21: -0.0240 S22: -0.6577 S23: -0.1362 REMARK 3 S31: -0.9135 S32: -0.6437 S33: 0.0334 REMARK 3 TLS GROUP : 4 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 75 THROUGH 115 ) REMARK 3 ORIGIN FOR THE GROUP (A): 7.3038 -4.9995 -6.3690 REMARK 3 T TENSOR REMARK 3 T11: 0.4140 T22: 0.5811 REMARK 3 T33: 0.8007 T12: 0.0801 REMARK 3 T13: -0.1134 T23: -0.1501 REMARK 3 L TENSOR REMARK 3 L11: 5.3941 L22: 3.2292 REMARK 3 L33: 4.7705 L12: -0.6269 REMARK 3 L13: 0.8770 L23: 0.9380 REMARK 3 S TENSOR REMARK 3 S11: -0.6112 S12: -0.3362 S13: 1.8926 REMARK 3 S21: 0.1826 S22: -0.0875 S23: 0.0660 REMARK 3 S31: -0.1913 S32: -0.8573 S33: 0.4525 REMARK 3 TLS GROUP : 5 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 116 THROUGH 151 ) REMARK 3 ORIGIN FOR THE GROUP (A): 16.3018 -28.6158 -19.5502 REMARK 3 T TENSOR REMARK 3 T11: 0.5542 T22: 0.6565 REMARK 3 T33: 0.3964 T12: -0.0433 REMARK 3 T13: -0.1876 T23: 0.1405 REMARK 3 L TENSOR REMARK 3 L11: 4.5692 L22: 5.5582 REMARK 3 L33: 4.9101 L12: -1.9613 REMARK 3 L13: 1.6180 L23: -2.3271 REMARK 3 S TENSOR REMARK 3 S11: 0.6504 S12: 0.6011 S13: -0.5477 REMARK 3 S21: -0.7750 S22: -0.0797 S23: 0.0581 REMARK 3 S31: 1.0019 S32: -0.1071 S33: -0.5841 REMARK 3 TLS GROUP : 6 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 152 THROUGH 199 ) REMARK 3 ORIGIN FOR THE GROUP (A): 16.4055 -31.5727 -11.4675 REMARK 3 T TENSOR REMARK 3 T11: 0.6107 T22: 0.5791 REMARK 3 T33: 0.4148 T12: -0.0826 REMARK 3 T13: -0.1236 T23: 0.2259 REMARK 3 L TENSOR REMARK 3 L11: 3.8495 L22: 4.4175 REMARK 3 L33: 3.4220 L12: -0.3732 REMARK 3 L13: 0.3345 L23: -0.6705 REMARK 3 S TENSOR REMARK 3 S11: 0.4045 S12: -0.1444 S13: -0.5071 REMARK 3 S21: 0.4045 S22: 0.0892 S23: 0.2176 REMARK 3 S31: 0.7817 S32: -0.4787 S33: -0.4275 REMARK 3 TLS GROUP : 7 REMARK 3 SELECTION: CHAIN 'H' AND (RESID 200 THROUGH 219 ) REMARK 3 ORIGIN FOR THE GROUP (A): 8.8631 -32.6765 -16.0648 REMARK 3 T TENSOR REMARK 3 T11: 0.6075 T22: 0.6993 REMARK 3 T33: 0.6117 T12: -0.1764 REMARK 3 T13: -0.1695 T23: 0.2394 REMARK 3 L TENSOR REMARK 3 L11: 6.4416 L22: 3.1179 REMARK 3 L33: 5.8582 L12: -2.3623 REMARK 3 L13: 1.7624 L23: -4.0702 REMARK 3 S TENSOR REMARK 3 S11: 0.5204 S12: -0.3877 S13: -1.3534 REMARK 3 S21: -0.2611 S22: 0.4261 S23: 1.4564 REMARK 3 S31: 0.9459 S32: -0.9512 S33: -0.8270 REMARK 3 TLS GROUP : 8 REMARK 3 SELECTION: CHAIN 'L' AND (RESID 1 THROUGH 38 ) REMARK 3 ORIGIN FOR THE GROUP (A): 25.3814 -3.6714 7.8748 REMARK 3 T TENSOR REMARK 3 T11: 0.6933 T22: 0.6595 REMARK 3 T33: 1.4151 T12: 0.2700 REMARK 3 T13: -0.5577 T23: -0.7252 REMARK 3 L TENSOR REMARK 3 L11: 1.0242 L22: 0.4086 REMARK 3 L33: 1.3229 L12: 0.3065 REMARK 3 L13: -0.1953 L23: 0.6602 REMARK 3 S TENSOR REMARK 3 S11: -0.6987 S12: -1.0042 S13: 2.0276 REMARK 3 S21: 0.8019 S22: -0.2635 S23: -0.9861 REMARK 3 S31: -0.5932 S32: 0.3903 S33: -0.6553 REMARK 3 TLS GROUP : 9 REMARK 3 SELECTION: CHAIN 'L' AND (RESID 39 THROUGH 101 ) REMARK 3 ORIGIN FOR THE GROUP (A): 19.1850 -7.1380 9.4870 REMARK 3 T TENSOR REMARK 3 T11: 0.6899 T22: 0.7239 REMARK 3 T33: 1.0495 T12: 0.3687 REMARK 3 T13: -0.3418 T23: -0.5973 REMARK 3 L TENSOR REMARK 3 L11: 1.2240 L22: 0.8295 REMARK 3 L33: 3.0707 L12: 0.3658 REMARK 3 L13: 0.2813 L23: 0.3740 REMARK 3 S TENSOR REMARK 3 S11: -0.6873 S12: -1.2620 S13: 1.8609 REMARK 3 S21: 0.4993 S22: 0.5879 S23: -0.3409 REMARK 3 S31: 0.0293 S32: -0.5344 S33: 0.2818 REMARK 3 TLS GROUP : 10 REMARK 3 SELECTION: CHAIN 'L' AND (RESID 102 THROUGH 113 ) REMARK 3 ORIGIN FOR THE GROUP (A): 31.5017 -24.2530 4.6763 REMARK 3 T TENSOR REMARK 3 T11: 0.7587 T22: 0.6370 REMARK 3 T33: 0.2890 T12: 0.3265 REMARK 3 T13: -0.4199 T23: -0.2215 REMARK 3 L TENSOR REMARK 3 L11: 8.8522 L22: 6.6092 REMARK 3 L33: 1.5177 L12: -2.1665 REMARK 3 L13: -2.9639 L23: 2.4914 REMARK 3 S TENSOR REMARK 3 S11: 0.5464 S12: -0.4858 S13: 0.8351 REMARK 3 S21: -0.6563 S22: -0.0888 S23: 0.0648 REMARK 3 S31: 0.4717 S32: 0.3210 S33: 0.7534 REMARK 3 TLS GROUP : 11 REMARK 3 SELECTION: CHAIN 'L' AND (RESID 114 THROUGH 150 ) REMARK 3 ORIGIN FOR THE GROUP (A): 28.9853 -34.3464 -14.4489 REMARK 3 T TENSOR REMARK 3 T11: 0.6658 T22: 0.4500 REMARK 3 T33: 0.4102 T12: 0.1021 REMARK 3 T13: -0.2504 T23: 0.0112 REMARK 3 L TENSOR REMARK 3 L11: 1.7551 L22: 3.3497 REMARK 3 L33: 6.0565 L12: -0.7004 REMARK 3 L13: -1.0609 L23: 1.7403 REMARK 3 S TENSOR REMARK 3 S11: 0.1500 S12: 0.2017 S13: -0.0614 REMARK 3 S21: 0.0544 S22: 0.2631 S23: -0.0913 REMARK 3 S31: 0.8343 S32: -0.1032 S33: -0.3995 REMARK 3 TLS GROUP : 12 REMARK 3 SELECTION: CHAIN 'L' AND (RESID 151 THROUGH 163 ) REMARK 3 ORIGIN FOR THE GROUP (A): 36.0630 -30.6369 -19.3957 REMARK 3 T TENSOR REMARK 3 T11: 0.5308 T22: 0.6209 REMARK 3 T33: 0.5052 T12: 0.1174 REMARK 3 T13: -0.1315 T23: -0.0614 REMARK 3 L TENSOR REMARK 3 L11: 4.4267 L22: 4.7171 REMARK 3 L33: 3.8390 L12: 0.0313 REMARK 3 L13: -2.5386 L23: 3.3267 REMARK 3 S TENSOR REMARK 3 S11: -0.0842 S12: -0.3585 S13: 0.0548 REMARK 3 S21: 0.3308 S22: 0.7142 S23: -0.6993 REMARK 3 S31: 1.0201 S32: 0.9660 S33: -0.7387 REMARK 3 TLS GROUP : 13 REMARK 3 SELECTION: CHAIN 'L' AND (RESID 164 THROUGH 213 ) REMARK 3 ORIGIN FOR THE GROUP (A): 31.9370 -35.5453 -13.7535 REMARK 3 T TENSOR REMARK 3 T11: 0.7405 T22: 0.5434 REMARK 3 T33: 0.5376 T12: 0.1267 REMARK 3 T13: -0.3001 T23: -0.0695 REMARK 3 L TENSOR REMARK 3 L11: 1.9955 L22: 0.9208 REMARK 3 L33: 3.7613 L12: -0.6046 REMARK 3 L13: -0.2484 L23: -0.4732 REMARK 3 S TENSOR REMARK 3 S11: 0.1385 S12: 0.1358 S13: 0.0859 REMARK 3 S21: 0.1328 S22: 0.5483 S23: -0.1613 REMARK 3 S31: 1.2368 S32: 0.1678 S33: -0.5897 REMARK 3 REMARK 3 NCS DETAILS REMARK 3 NUMBER OF NCS GROUPS : NULL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL REMARK 4 REMARK 4 9MFN COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB. REMARK 100 THE DEPOSITION ID IS D_1000290850. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 12-MAY-21 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 8.5 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : DIAMOND REMARK 200 BEAMLINE : I04 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.9795 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS EIGER2 XE 16M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : XDS REMARK 200 DATA SCALING SOFTWARE : AIMLESS REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 208056 REMARK 200 RESOLUTION RANGE HIGH (A) : 2.350 REMARK 200 RESOLUTION RANGE LOW (A) : 96.790 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 100.0 REMARK 200 DATA REDUNDANCY : 10.70 REMARK 200 R MERGE (I) : NULL REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 7.9000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.35 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.43 REMARK 200 COMPLETENESS FOR SHELL (%) : 100.0 REMARK 200 DATA REDUNDANCY IN SHELL : 10.10 REMARK 200 R MERGE FOR SHELL (I) : NULL REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 FOR SHELL : 1.000 REMARK 200 REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT REMARK 200 SOFTWARE USED: PHASER REMARK 200 STARTING MODEL: NULL REMARK 200 REMARK 200 REMARK: NEEDLE-LIKE CRYSTALS REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 51.82 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.55 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: ADI-64596 (HUMAN FAB) WAS RECEIVED AT REMARK 280 A CONCENTRATION OF 11.35 MG/ML IN A BUFFER CONTAINING 2 MM TRIS- REMARK 280 HCL PH 8.0 AND 150 MM NACL. THE PACT, BCS, AND JCSG+ SCREENS REMARK 280 (ALL FROM MOLECULAR DIMENSIONS LTD.) WERE INITIALLY SET UP USING REMARK 280 A MOSQUITO CRYSTALLIZATION ROBOT (STP LABTECH). SINCE CRYSTALS REMARK 280 OBTAINED FROM THESE INITIAL SCREENS ONLY GAVE RISE TO LOW- REMARK 280 RESOLUTION X-RAY DIFFRACTION, CRYSTAL SEED SOLUTIONS WERE REMARK 280 PREPARED AND APPLIED IN THE SETUP OF THE BCS, PACT, AND ADDITIVE REMARK 280 SCREENS (HAMPTON RESEARCH). SITTING DROPS OF 160 NL PROTEIN AND REMARK 280 160 NL PRECIPITANT SOLUTION WERE LEFT TO EQUILIBRATE AGAINST A REMARK 280 40 UL RESERVOIR AT 20 DEGREES C. AFTER A FEW DAYS, PLATE- AND REMARK 280 NEEDLE-LIKE CRYSTALS APPEARED IN SEVERAL CONDITIONS. THE REMARK 280 PRECIPITANT SOLUTION GIVING RISE TO THE BEST-DIFFRACTING CRYSTAL REMARK 280 CONTAINED 75 MM TRIS PH 8.5, 25 MM BIS-TRIS-PROPANE PH 8.5, 22.5% REMARK 280 (V/V) PEG SMEAR LOW, 5% (W/V) PEG3350, AND 50 MM NABR, VAPOR REMARK 280 DIFFUSION, SITTING DROP, TEMPERATURE 293K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 31 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -Y,X-Y,Z+1/3 REMARK 290 3555 -X+Y,-X,Z+2/3 REMARK 290 REMARK 290 WHERE NNN -> OPERATOR NUMBER REMARK 290 MMM -> TRANSLATION VECTOR REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS REMARK 290 THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM REMARK 290 RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY REMARK 290 RELATED MOLECULES. REMARK 290 SMTRY1 1 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 1 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY3 1 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 2 -0.500000 -0.866025 0.000000 0.00000 REMARK 290 SMTRY2 2 0.866025 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 32.26500 REMARK 290 SMTRY1 3 -0.500000 0.866025 0.000000 0.00000 REMARK 290 SMTRY2 3 -0.866025 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 64.53000 REMARK 290 REMARK 290 REMARK: NULL REMARK 300 REMARK 300 BIOMOLECULE: 1 REMARK 300 SEE REMARK 350 FOR THE AUTHOR PROVIDED AND/OR PROGRAM REMARK 300 GENERATED ASSEMBLY INFORMATION FOR THE STRUCTURE IN REMARK 300 THIS ENTRY. THE REMARK MAY ALSO PROVIDE INFORMATION ON REMARK 300 BURIED SURFACE AREA. REMARK 350 REMARK 350 COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN REMARK 350 BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE REMARK 350 MOLECULE CAN BE GENERATED BY APPLYING BIOMT TRANSFORMATIONS REMARK 350 GIVEN BELOW. BOTH NON-CRYSTALLOGRAPHIC AND REMARK 350 CRYSTALLOGRAPHIC OPERATIONS ARE GIVEN. REMARK 350 REMARK 350 BIOMOLECULE: 1 REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: DIMERIC REMARK 350 SOFTWARE DETERMINED QUATERNARY STRUCTURE: DIMERIC REMARK 350 SOFTWARE USED: PISA REMARK 350 TOTAL BURIED SURFACE AREA: 4190 ANGSTROM**2 REMARK 350 SURFACE AREA OF THE COMPLEX: 18700 ANGSTROM**2 REMARK 350 CHANGE IN SOLVENT FREE ENERGY: -12.0 KCAL/MOL REMARK 350 APPLY THE FOLLOWING TO CHAINS: H, L REMARK 350 BIOMT1 1 1.000000 0.000000 0.000000 0.00000 REMARK 350 BIOMT2 1 0.000000 1.000000 0.000000 0.00000 REMARK 350 BIOMT3 1 0.000000 0.000000 1.000000 0.00000 REMARK 465 REMARK 465 MISSING RESIDUES REMARK 465 THE FOLLOWING RESIDUES WERE NOT LOCATED IN THE REMARK 465 EXPERIMENT. (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN REMARK 465 IDENTIFIER; SSSEQ=SEQUENCE NUMBER; I=INSERTION CODE.) REMARK 465 REMARK 465 M RES C SSSEQI REMARK 465 SER H 133 REMARK 465 SER H 134 REMARK 465 LYS H 135 REMARK 465 SER H 136 REMARK 465 THR H 137 REMARK 465 SER H 138 REMARK 465 GLY H 139 REMARK 465 GLY H 140 REMARK 465 LYS H 220 REMARK 465 SER H 221 REMARK 465 CYS H 222 REMARK 465 CYS L 214 REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: TORSION ANGLES REMARK 500 REMARK 500 TORSION ANGLES OUTSIDE THE EXPECTED RAMACHANDRAN REGIONS: REMARK 500 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; REMARK 500 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE). REMARK 500 REMARK 500 STANDARD TABLE: REMARK 500 FORMAT:(10X,I3,1X,A3,1X,A1,I4,A1,4X,F7.2,3X,F7.2) REMARK 500 REMARK 500 EXPECTED VALUES: GJ KLEYWEGT AND TA JONES (1996). PHI/PSI- REMARK 500 CHOLOGY: RAMACHANDRAN REVISITED. STRUCTURE 4, 1395 - 1400 REMARK 500 REMARK 500 M RES CSSEQI PSI PHI REMARK 500 SER H 15 -4.02 67.42 REMARK 500 SER H 30 47.34 -91.02 REMARK 500 LYS H 45 17.84 -141.80 REMARK 500 ASN H 60 -55.50 -151.42 REMARK 500 TYR H 61 149.39 69.63 REMARK 500 SER L 30 -119.99 54.14 REMARK 500 ALA L 51 -21.87 68.42 REMARK 500 SER L 52 3.74 -150.91 REMARK 500 ASN L 138 64.24 61.74 REMARK 500 REMARK 500 REMARK: NULL DBREF 9MFN H 1 222 PDB 9MFN 9MFN 1 222 DBREF 9MFN L 1 214 PDB 9MFN 9MFN 1 214 SEQRES 1 H 222 PCA VAL GLN LEU GLN GLU SER GLY PRO GLY LEU VAL LYS SEQRES 2 H 222 PRO SER GLU THR LEU SER LEU THR CYS THR VAL SER GLY SEQRES 3 H 222 GLY SER VAL SER SER GLY ASP TYR TYR TRP THR TRP ILE SEQRES 4 H 222 ARG GLN SER PRO GLY LYS GLY LEU GLU TRP ILE GLY HIS SEQRES 5 H 222 ILE TYR TYR SER GLY ASN THR ASN TYR ASN PRO SER LEU SEQRES 6 H 222 LYS SER ARG LEU THR ILE SER ILE ASP THR SER LYS THR SEQRES 7 H 222 GLN PHE SER LEU LYS LEU SER SER VAL THR ALA ALA ASP SEQRES 8 H 222 THR ALA ILE TYR TYR CYS VAL ARG ASP ARG VAL THR GLY SEQRES 9 H 222 ALA PHE ASP ILE TRP GLY GLN GLY THR MET VAL THR VAL SEQRES 10 H 222 SER SER ALA SER THR LYS GLY PRO SER VAL PHE PRO LEU SEQRES 11 H 222 ALA PRO SER SER LYS SER THR SER GLY GLY THR ALA ALA SEQRES 12 H 222 LEU GLY CYS GLN VAL GLU ASP TYR PHE PRO GLU PRO VAL SEQRES 13 H 222 THR VAL SER TRP ASN SER GLY ALA LEU THR SER GLY VAL SEQRES 14 H 222 HIS THR PHE PRO ALA VAL LEU GLN SER SER GLY LEU TYR SEQRES 15 H 222 GLU LEU SER SER VAL VAL THR VAL PRO SER SER SER LEU SEQRES 16 H 222 GLY THR GLN THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SEQRES 17 H 222 SER ASN THR LYS VAL ASP LYS LYS VAL GLU PRO LYS SER SEQRES 18 H 222 CYS SEQRES 1 L 214 ASP ILE GLN MET THR GLN SER PRO SER SER LEU SER ALA SEQRES 2 L 214 SER VAL GLY ASP ARG VAL THR ILE THR CYS GLN ALA SER SEQRES 3 L 214 GLN ASP ILE SER ASN TYR LEU ASN TRP TYR GLN GLN LYS SEQRES 4 L 214 PRO GLY LYS ALA PRO LYS LEU LEU ILE TYR ASP ALA SER SEQRES 5 L 214 ASN LEU GLU THR GLY VAL PRO SER ARG PHE SER GLY SER SEQRES 6 L 214 GLY SER GLY THR ASP PHE THR PHE THR ILE SER SER LEU SEQRES 7 L 214 GLN PRO GLU ASP ILE ALA THR TYR PHE CYS GLN HIS PHE SEQRES 8 L 214 ASP HIS LEU PRO LEU ALA PHE GLY GLY GLY THR LYS VAL SEQRES 9 L 214 GLU ILE LYS ARG THR VAL ALA ALA PRO SER VAL PHE ILE SEQRES 10 L 214 PHE PRO PRO SER ASP GLU GLN LEU LYS SER GLY ARG ALA SEQRES 11 L 214 SER VAL VAL CYS LEU LEU ASN ASN PHE TYR PRO ARG GLU SEQRES 12 L 214 ALA LYS VAL GLN TRP LYS VAL ASP ASN ALA LEU GLN SER SEQRES 13 L 214 GLY ASN SER GLN GLU SER VAL THR GLU GLN ASP SER LYS SEQRES 14 L 214 ASP SER THR TYR SER LEU SER SER ARG LEU GLN LEU SER SEQRES 15 L 214 LYS ALA ASP TYR GLU LYS HIS LYS VAL TYR ALA CYS GLU SEQRES 16 L 214 VAL THR HIS GLN GLY LEU SER SER PRO VAL THR LYS SER SEQRES 17 L 214 PHE ASN ARG GLY GLU CYS HET PCA H 1 8 HET BR H 301 1 HET BR H 302 1 HET PEG H 303 7 HET BR L 301 1 HET PEG L 302 7 HETNAM PCA PYROGLUTAMIC ACID HETNAM BR BROMIDE ION HETNAM PEG DI(HYDROXYETHYL)ETHER FORMUL 1 PCA C5 H7 N O3 FORMUL 3 BR 3(BR 1-) FORMUL 5 PEG 2(C4 H10 O3) FORMUL 8 HOH *38(H2 O) HELIX 1 AA1 ASN H 62 LYS H 66 5 5 HELIX 2 AA2 THR H 88 THR H 92 5 5 HELIX 3 AA3 SER H 162 ALA H 164 5 3 HELIX 4 AA4 PRO H 191 THR H 197 5 7 HELIX 5 AA5 LYS H 207 ASN H 210 5 4 HELIX 6 AA6 GLN L 79 ILE L 83 5 5 HELIX 7 AA7 SER L 121 SER L 127 1 7 HELIX 8 AA8 LYS L 183 GLU L 187 1 5 SHEET 1 AA1 4 GLN H 3 SER H 7 0 SHEET 2 AA1 4 LEU H 18 SER H 25 -1 O THR H 23 N GLN H 5 SHEET 3 AA1 4 GLN H 79 LEU H 84 -1 O LEU H 84 N LEU H 18 SHEET 4 AA1 4 LEU H 69 ASP H 74 -1 N ASP H 74 O GLN H 79 SHEET 1 AA2 5 LEU H 11 VAL H 12 0 SHEET 2 AA2 5 THR H 113 VAL H 117 1 O THR H 116 N VAL H 12 SHEET 3 AA2 5 ALA H 93 ARG H 101 -1 N TYR H 95 O THR H 113 SHEET 4 AA2 5 TYR H 34 SER H 42 -1 N ILE H 39 O TYR H 96 SHEET 5 AA2 5 GLY H 46 ILE H 53 -1 O GLU H 48 N ARG H 40 SHEET 1 AA3 4 LEU H 11 VAL H 12 0 SHEET 2 AA3 4 THR H 113 VAL H 117 1 O THR H 116 N VAL H 12 SHEET 3 AA3 4 ALA H 93 ARG H 101 -1 N TYR H 95 O THR H 113 SHEET 4 AA3 4 PHE H 106 TRP H 109 -1 O ILE H 108 N ARG H 99 SHEET 1 AA4 4 SER H 126 LEU H 130 0 SHEET 2 AA4 4 ALA H 142 TYR H 151 -1 O GLN H 147 N PHE H 128 SHEET 3 AA4 4 TYR H 182 VAL H 190 -1 O LEU H 184 N VAL H 148 SHEET 4 AA4 4 VAL H 169 THR H 171 -1 N HIS H 170 O VAL H 187 SHEET 1 AA5 4 SER H 126 LEU H 130 0 SHEET 2 AA5 4 ALA H 142 TYR H 151 -1 O GLN H 147 N PHE H 128 SHEET 3 AA5 4 TYR H 182 VAL H 190 -1 O LEU H 184 N VAL H 148 SHEET 4 AA5 4 VAL H 175 LEU H 176 -1 N VAL H 175 O GLU H 183 SHEET 1 AA6 3 THR H 157 TRP H 160 0 SHEET 2 AA6 3 TYR H 200 HIS H 206 -1 O ASN H 203 N SER H 159 SHEET 3 AA6 3 THR H 211 VAL H 217 -1 O THR H 211 N HIS H 206 SHEET 1 AA7 4 MET L 4 SER L 7 0 SHEET 2 AA7 4 VAL L 19 ALA L 25 -1 O GLN L 24 N THR L 5 SHEET 3 AA7 4 ASP L 70 ILE L 75 -1 O PHE L 73 N ILE L 21 SHEET 4 AA7 4 PHE L 62 SER L 67 -1 N SER L 63 O THR L 74 SHEET 1 AA8 6 SER L 10 SER L 14 0 SHEET 2 AA8 6 THR L 102 LYS L 107 1 O GLU L 105 N LEU L 11 SHEET 3 AA8 6 ALA L 84 HIS L 90 -1 N ALA L 84 O VAL L 104 SHEET 4 AA8 6 LEU L 33 GLN L 38 -1 N TYR L 36 O PHE L 87 SHEET 5 AA8 6 LYS L 45 TYR L 49 -1 O ILE L 48 N TRP L 35 SHEET 6 AA8 6 ASN L 53 LEU L 54 -1 O ASN L 53 N TYR L 49 SHEET 1 AA9 4 SER L 10 SER L 14 0 SHEET 2 AA9 4 THR L 102 LYS L 107 1 O GLU L 105 N LEU L 11 SHEET 3 AA9 4 ALA L 84 HIS L 90 -1 N ALA L 84 O VAL L 104 SHEET 4 AA9 4 ALA L 97 PHE L 98 -1 O ALA L 97 N HIS L 90 SHEET 1 AB1 4 SER L 114 PHE L 118 0 SHEET 2 AB1 4 ARG L 129 PHE L 139 -1 O LEU L 135 N PHE L 116 SHEET 3 AB1 4 TYR L 173 SER L 182 -1 O SER L 177 N CYS L 134 SHEET 4 AB1 4 SER L 159 VAL L 163 -1 N SER L 162 O SER L 176 SHEET 1 AB2 3 LYS L 145 VAL L 150 0 SHEET 2 AB2 3 VAL L 191 THR L 197 -1 O ALA L 193 N LYS L 149 SHEET 3 AB2 3 VAL L 205 ASN L 210 -1 O LYS L 207 N CYS L 194 SSBOND 1 CYS H 22 CYS H 97 1555 1555 2.05 SSBOND 2 CYS H 146 CYS H 202 1555 1555 2.03 SSBOND 3 CYS L 23 CYS L 88 1555 1555 2.04 SSBOND 4 CYS L 134 CYS L 194 1555 1555 2.03 LINK C PCA H 1 N VAL H 2 1555 1555 1.33 CISPEP 1 PHE H 152 PRO H 153 0 -1.18 CISPEP 2 GLU H 154 PRO H 155 0 0.21 CISPEP 3 SER L 7 PRO L 8 0 -5.60 CISPEP 4 LEU L 94 PRO L 95 0 0.88 CISPEP 5 TYR L 140 PRO L 141 0 5.39 CRYST1 65.594 65.594 96.795 90.00 90.00 120.00 P 31 3 ORIGX1 1.000000 0.000000 0.000000 0.00000 ORIGX2 0.000000 1.000000 0.000000 0.00000 ORIGX3 0.000000 0.000000 1.000000 0.00000 SCALE1 0.015245 0.008802 0.000000 0.00000 SCALE2 0.000000 0.017604 0.000000 0.00000 SCALE3 0.000000 0.000000 0.010331 0.00000