HEADER BLOOD CLOTTING 22-DEC-99 1C5N TITLE STRUCTURAL BASIS FOR SELECTIVITY OF A SMALL MOLECULE, S1- TITLE 2 BINDING, SUB-MICROMOLAR INHIBITOR OF UROKINASE TYPE TITLE 3 PLASMINOGEN ACTIVATOR COMPND MOL_ID: 1; COMPND 2 MOLECULE: PROTEIN (ALPHA THROMBIN (LIGHT CHAIN)); COMPND 3 CHAIN: L; COMPND 4 EC: 3.4.21.5; COMPND 5 MOL_ID: 2; COMPND 6 MOLECULE: PROTEIN (ALPHA THROMBIN (HEAVY CHAIN)); COMPND 7 CHAIN: H; COMPND 8 EC: 3.4.21.5; COMPND 9 MOL_ID: 3; COMPND 10 MOLECULE: PROTEIN (ACETYL HIRUDIN); COMPND 11 CHAIN: I; COMPND 12 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 3 ORGANISM_COMMON: HUMAN; SOURCE 4 MOL_ID: 2; SOURCE 5 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 6 ORGANISM_COMMON: HUMAN; SOURCE 7 MOL_ID: 3; SOURCE 8 SYNTHETIC: YES KEYWDS SELECTIVE, S1 SITE INHIBITOR, STRUCTURE-BASED DRUG DESIGN, KEYWDS 2 UROKINASE, TRYPSIN, THROMBIN EXPDTA X-RAY DIFFRACTION AUTHOR B.A.KATZ,R.MACKMAN,C.LUONG,K.RADIKA,A.MARTELLI, AUTHOR 2 P.A.SPRENGELER,J.WANG,H.CHAN,L.WONG REVDAT 3 06-JUL-04 1C5N 3 JRNL REMARK ATOM FORMUL REVDAT 3 2 3 MASTER REVDAT 2 26-SEP-01 1C5N 3 ATOM REVDAT 1 22-DEC-00 1C5N 0 JRNL AUTH B.A.KATZ,R.MACKMAN,C.LUONG,K.RADIKA,A.MARTELLI, JRNL AUTH 2 P.A.SPRENGELER,J.WANG,H.CHAN,L.WONG JRNL TITL STRUCTURAL BASIS FOR SELECTIVITY OF A SMALL JRNL TITL 2 MOLECULE, S1-BINDING, SUBMICROMOLAR INHIBITOR OF JRNL TITL 3 UROKINASE-TYPE PLASMINOGEN ACTIVATOR. JRNL REF CHEM.BIOL. V. 7 299 2000 JRNL REFN ASTM CBOLE2 UK ISSN 1074-5521 REMARK 1 REMARK 2 REMARK 2 RESOLUTION. 1.50 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : X-PLOR 3.1 REMARK 3 AUTHORS : BRUNGER REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.50 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 7.50 REMARK 3 DATA CUTOFF (SIGMA(F)) : 2.000 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : NULL REMARK 3 DATA CUTOFF LOW (ABS(F)) : NULL REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 65.0 REMARK 3 NUMBER OF REFLECTIONS : 37810 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 CROSS-VALIDATION METHOD : X-PLOR REMARK 3 FREE R VALUE TEST SET SELECTION : NULL REMARK 3 R VALUE (WORKING SET) : 0.220 REMARK 3 FREE R VALUE : 0.246 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 10.000 REMARK 3 FREE R VALUE TEST SET COUNT : 3800 REMARK 3 ESTIMATED ERROR OF FREE R VALUE : NULL REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN. REMARK 3 TOTAL NUMBER OF BINS USED : 8 REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 1.50 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 1.57 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 36.10 REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 2331 REMARK 3 BIN R VALUE (WORKING SET) : 0.4980 REMARK 3 BIN FREE R VALUE : 0.5080 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : NULL REMARK 3 BIN FREE R VALUE TEST SET COUNT : 282 REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : NULL REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 4831 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 23 REMARK 3 SOLVENT ATOMS : 765 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 ESTIMATED COORDINATE ERROR. REMARK 3 ESD FROM LUZZATI PLOT (A) : NULL REMARK 3 ESD FROM SIGMAA (A) : NULL REMARK 3 LOW RESOLUTION CUTOFF (A) : NULL REMARK 3 REMARK 3 CROSS-VALIDATED ESTIMATED COORDINATE ERROR. REMARK 3 ESD FROM C-V LUZZATI PLOT (A) : NULL REMARK 3 ESD FROM C-V SIGMAA (A) : NULL REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES. REMARK 3 BOND LENGTHS (A) : 0.018 REMARK 3 BOND ANGLES (DEGREES) : 4.00 REMARK 3 DIHEDRAL ANGLES (DEGREES) : 26.10 REMARK 3 IMPROPER ANGLES (DEGREES) : 0.49 REMARK 3 REMARK 3 ISOTROPIC THERMAL MODEL : NULL REMARK 3 REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA REMARK 3 MAIN-CHAIN BOND (A**2) : NULL ; NULL REMARK 3 MAIN-CHAIN ANGLE (A**2) : NULL ; NULL REMARK 3 SIDE-CHAIN BOND (A**2) : NULL ; NULL REMARK 3 SIDE-CHAIN ANGLE (A**2) : NULL ; NULL REMARK 3 REMARK 3 NCS MODEL : NULL REMARK 3 REMARK 3 NCS RESTRAINTS. RMS SIGMA/WEIGHT REMARK 3 GROUP 1 POSITIONAL (A) : NULL ; NULL REMARK 3 GROUP 1 B-FACTOR (A**2) : NULL ; NULL REMARK 3 REMARK 3 PARAMETER FILE 1 : TH6860_PARMALLH3X.PRO REMARK 3 PARAMETER FILE 2 : TH6860_PARAM11_UCSF.WAT REMARK 3 PARAMETER FILE 3 : NULL REMARK 3 TOPOLOGY FILE 1 : TH6860_TOPALLH6X.PRO REMARK 3 TOPOLOGY FILE 2 : NULL REMARK 3 TOPOLOGY FILE 3 : NULL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: BULK SOLVENT TERMS INCLUDED IN FOB REMARK 3 FILE CREATED WITH STANDARD X-PLOR SCRIPT. MET_H84, GLU_H97A REMARK 3 MET_H106, AND MET_H210 WAS SIMULTANEOUSLY REFINED IN TWO REMARK 3 CONFORMATIONS. NO DENSITY WAS OBSERVED FOR TRP148, THR149, REMARK 3 ALA149A, ASN149B, VAL149C, GLY149D, AND LYS149E IN THE REMARK 3 AUTOLYSIS LOOP, AND THESE RESIDUES ARE NOT INCLUDED IN THE REMARK 3 MODEL. DISORDERED WATERS INCLUDE: HOH395 WHICH IS IN A SPECIAL REMARK 3 POSITION. HOH396 IS CLOSE TO A SYMMETRY RELATED EQUIVALENT OF REMARK 3 ITSELF; HOH422 WHICH IS CLOSE TO HOH432; HOH434 IS CLOSE TO A REMARK 3 SYMMETRY RELATED EQUIVALENT OF ITSELF; (THE ABOVE WATERS HAVE REMARK 3 VERY STRONG DENSITY AND THEIR REFINED OCCUPANCIES ARE REMARK 3 SIGNIFICANTLY GREATER THAN UNITY. THEY MAY REFLECT A DIFFERENT REMARK 3 FEATURE THAN DISORDERED WATERS); HOH663 WHICH IS CLOSE TO A REMARK 3 SYMMETRY-RELATED EQUIVALENT OF HOH575 AND TO A SYMMETRY- REMARK 3 RELATED EQUIVALENT OF RESIDUE L1E; HOH511 WHICH IS CLOSE TO REMARK 3 HOH758; HOH718 WHICH IS CLOSE TO HOH720; HOH586 WHICH IS CLOSE REMARK 3 TO HOH721. HIS_H57 IS MONOPROTONATED ON THE DELTA NITROGEN. REMARK 3 HIS_H91 AND HIS_H119 ARE MONOPROTONATED ON THE EPSILON NITROGEN REMARK 4 REMARK 4 1C5N COMPLIES WITH FORMAT V. 3.0, 1-DEC-2006 REMARK 4 REMARK 4 THIS IS THE REMEDIATED VERSION OF THIS PDB ENTRY. REMARK 4 REMEDIATED DATA FILE REVISION 3.101 (2007-05-29) REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB. REMARK 100 THE RCSB ID CODE IS RCSB001367. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 08-APR-1999 REMARK 200 TEMPERATURE (KELVIN) : 298.0 REMARK 200 PH : 7.50 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : N REMARK 200 RADIATION SOURCE : NULL REMARK 200 BEAMLINE : NULL REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 1.54 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : MSC MIRRORS REMARK 200 REMARK 200 DETECTOR TYPE : ROTATING ANODE REMARK 200 DETECTOR MANUFACTURER : RIGAKU RAXIS IV++ REMARK 200 INTENSITY-INTEGRATION SOFTWARE : BIOTEX (MSC) REMARK 200 DATA SCALING SOFTWARE : BIOTEX REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : NULL REMARK 200 RESOLUTION RANGE HIGH (A) : 1.330 REMARK 200 RESOLUTION RANGE LOW (A) : 36.130 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 1.000 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 65.0 REMARK 200 DATA REDUNDANCY : 2.100 REMARK 200 R MERGE (I) : 0.06300 REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : 9.0000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.50 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.57 REMARK 200 COMPLETENESS FOR SHELL (%) : 36.1 REMARK 200 DATA REDUNDANCY IN SHELL : NULL REMARK 200 R MERGE FOR SHELL (I) : 0.24500 REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 FOR SHELL : 1.900 REMARK 200 REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: DIFFERENCE FOURIER PLUS REMARK 200 REFINEMENT REMARK 200 SOFTWARE USED: X-PLOR, QUANTA, INSIGHTII REMARK 200 STARTING MODEL: NULL REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 36.70 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.65 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: THROMBIN WAS PURCHASED FROM REMARK 280 HAEMATOLOGIC TECHNOLOGIES, INC. AND ACETYL-HIRUDIN FROM REMARK 280 BACHEM. THROMBIN WAS PREPARED AS DESCRIBED (SKRZPCZAK-JANKUN REMARK 280 ET AL., 1991). THROMBIN (1.0 MG/ML IN 50 MM HEPES, 50 % REMARK 280 GLYCEROL, PH 7.0) WAS INCUBATED WITH 1.0 MM ACETYL-HIRUDIN, 10 REMARK 280 MM 4-IODOBENZO[B]THIOPHENE-2-CARBOXAMIDINE FOR 1 HR AT 4 DEG REMARK 280 C. GLYCEROL WAS REMOVED ANDTHE COMPLEX CONCENTRATED WITH A REMARK 280 CENTRICON 10 (AMICON) TO ABOUT 10 MG/ML AS DETERMINED BY THE REMARK 280 BIORAD PROTEIN ASSAY KIT USING BOVINE SERUM ALBUMIN. CRYSTALS REMARK 280 OF THROMBIN-ACETYL-HIRUDIN-4-IODOBENZO[B]THIOPHENE-2- REMARK 280 CARBOXAMIDINE WERE GROWN IN HANGING DROPS BY VAPOR DIFFUSION REMARK 280 AFTER STREAK SEEDING. THE DROPS WERE MADE FROM 5 MICROLITERS REMARK 280 OF COMPLEX AND 5 MICROLITERS OF RESERVOIR SOLUTION (0.10 M REMARK 280 TRIS,0.50 M NACL, 22 % (BY VOLUME) PEG 4K, PH 7.5)., VAPOR REMARK 280 DIFFUSION REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: C 1 2 1 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X,Y,-Z REMARK 290 3555 1/2+X,1/2+Y,Z REMARK 290 4555 1/2-X,1/2+Y,-Z 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 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 2 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 -1.000000 0.00000 REMARK 290 SMTRY1 3 1.000000 0.000000 0.000000 36.07000 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 36.13000 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 4 -1.000000 0.000000 0.000000 36.07000 REMARK 290 SMTRY2 4 0.000000 1.000000 0.000000 36.13000 REMARK 290 SMTRY3 4 0.000000 0.000000 -1.000000 0.00000 REMARK 290 REMARK 290 REMARK: NULL REMARK 300 REMARK 300 BIOMOLECULE: 1 REMARK 300 THIS ENTRY CONTAINS THE CRYSTALLOGRAPHIC ASYMMETRIC UNIT REMARK 300 WHICH CONSISTS OF 3 CHAIN(S). SEE REMARK 350 FOR REMARK 300 INFORMATION ON GENERATING THE BIOLOGICAL MOLECULE(S). REMARK 350 REMARK 350 GENERATING THE BIOMOLECULE 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 APPLY THE FOLLOWING TO CHAINS: L, H, I 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 TRP H 147A REMARK 465 THR H 147B REMARK 465 ALA H 147C REMARK 465 ASN H 147D REMARK 465 VAL H 147E REMARK 465 GLY H 147F REMARK 465 LYS H 147G REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: CLOSE CONTACTS IN SAME ASYMMETRIC UNIT REMARK 500 REMARK 500 THE FOLLOWING ATOMS ARE IN CLOSE CONTACT. REMARK 500 REMARK 500 ATM1 RES C SSEQI ATM2 RES C SSEQI REMARK 500 O SER L 1E O HOH 663 2.07 REMARK 500 O HOH 575 O HOH 663 2.08 REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: CLOSE CONTACTS REMARK 500 REMARK 500 THE FOLLOWING ATOMS THAT ARE RELATED BY CRYSTALLOGRAPHIC REMARK 500 SYMMETRY ARE IN CLOSE CONTACT. AN ATOM LOCATED WITHIN 0.15 REMARK 500 ANGSTROMS OF A SYMMETRY RELATED ATOM IS ASSUMED TO BE ON A REMARK 500 SPECIAL POSITION AND IS, THEREFORE, LISTED IN REMARK 375 REMARK 500 INSTEAD OF REMARK 500. ATOMS WITH NON-BLANK ALTERNATE REMARK 500 LOCATION INDICATORS ARE NOT INCLUDED IN THE CALCULATIONS. REMARK 500 REMARK 500 DISTANCE CUTOFF: REMARK 500 2.2 ANGSTROMS FOR CONTACTS NOT INVOLVING HYDROGEN ATOMS REMARK 500 1.6 ANGSTROMS FOR CONTACTS INVOLVING HYDROGEN ATOMS REMARK 500 REMARK 500 ATM1 RES C SSEQI ATM2 RES C SSEQI SSYMOP DISTANCE REMARK 500 O HOH 395 O HOH 395 2555 0.39 REMARK 500 O HOH 396 O HOH 396 2555 1.63 REMARK 500 O HOH 434 O HOH 434 2555 2.05 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 M RES CSSEQI PSI PHI REMARK 500 ASP L 14L 141.94 81.18 REMARK 525 REMARK 525 SOLVENT REMARK 525 THE FOLLOWING SOLVENT MOLECULES LIE FARTHER THAN EXPECTED REMARK 525 FROM THE PROTEIN OR NUCLEIC ACID MOLECULE AND MAY BE REMARK 525 ASSOCIATED WITH A SYMMETRY RELATED MOLECULE (M=MODEL REMARK 525 NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE REMARK 525 NUMBER; I=INSERTION CODE): REMARK 525 REMARK 525 M RES CSSEQI REMARK 525 HOH 389 DISTANCE = 5.55 ANGSTROMS REMARK 525 HOH 391 DISTANCE = 5.89 ANGSTROMS REMARK 525 HOH 552 DISTANCE = 5.45 ANGSTROMS REMARK 525 HOH 569 DISTANCE = 5.88 ANGSTROMS REMARK 525 HOH 615 DISTANCE = 6.37 ANGSTROMS REMARK 525 HOH 674 DISTANCE = 7.28 ANGSTROMS REMARK 525 HOH 675 DISTANCE = 7.81 ANGSTROMS REMARK 525 HOH 705 DISTANCE = 5.57 ANGSTROMS REMARK 525 HOH 725 DISTANCE = 5.20 ANGSTROMS REMARK 525 HOH 755 DISTANCE = 6.46 ANGSTROMS REMARK 525 HOH 757 DISTANCE = 7.63 ANGSTROMS DBREF 1C5N L 1H 15 UNP P00734 THRB_HUMAN 328 363 DBREF 1C5N H 16 247 UNP P00734 THRB_HUMAN 364 622 SEQRES 1 L 36 THR PHE GLY SER GLY GLU ALA ASP CYS GLY LEU ARG PRO SEQRES 2 L 36 LEU PHE GLU LYS LYS SER LEU GLU ASP LYS THR GLU ARG SEQRES 3 L 36 GLU LEU LEU GLU SER TYR ILE ASP GLY ARG SEQRES 1 H 259 ILE VAL GLU GLY SER ASP ALA GLU ILE GLY MET SER PRO SEQRES 2 H 259 TRP GLN VAL MET LEU PHE ARG LYS SER PRO GLN GLU LEU SEQRES 3 H 259 LEU CYS GLY ALA SER LEU ILE SER ASP ARG TRP VAL LEU SEQRES 4 H 259 THR ALA ALA HIS CYS LEU LEU TYR PRO PRO TRP ASP LYS SEQRES 5 H 259 ASN PHE THR GLU ASN ASP LEU LEU VAL ARG ILE GLY LYS SEQRES 6 H 259 HIS SER ARG THR ARG TYR GLU ARG ASN ILE GLU LYS ILE SEQRES 7 H 259 SER MET LEU GLU LYS ILE TYR ILE HIS PRO ARG TYR ASN SEQRES 8 H 259 TRP ARG GLU ASN LEU ASP ARG ASP ILE ALA LEU MET LYS SEQRES 9 H 259 LEU LYS LYS PRO VAL ALA PHE SER ASP TYR ILE HIS PRO SEQRES 10 H 259 VAL CYS LEU PRO ASP ARG GLU THR ALA ALA SER LEU LEU SEQRES 11 H 259 GLN ALA GLY TYR LYS GLY ARG VAL THR GLY TRP GLY ASN SEQRES 12 H 259 LEU LYS GLU THR TRP THR ALA ASN VAL GLY LYS GLY GLN SEQRES 13 H 259 PRO SER VAL LEU GLN VAL VAL ASN LEU PRO ILE VAL GLU SEQRES 14 H 259 ARG PRO VAL CYS LYS ASP SER THR ARG ILE ARG ILE THR SEQRES 15 H 259 ASP ASN MET PHE CYS ALA GLY TYR LYS PRO ASP GLU GLY SEQRES 16 H 259 LYS ARG GLY ASP ALA CYS GLU GLY ASP SER GLY GLY PRO SEQRES 17 H 259 PHE VAL MET LYS SER PRO PHE ASN ASN ARG TRP TYR GLN SEQRES 18 H 259 MET GLY ILE VAL SER TRP GLY GLU GLY CYS ASP ARG ASP SEQRES 19 H 259 GLY LYS TYR GLY PHE TYR THR HIS VAL PHE ARG LEU LYS SEQRES 20 H 259 LYS TRP ILE GLN LYS VAL ILE ASP GLN PHE GLY GLU SEQRES 1 I 10 ASP PHE GLU GLU ILE PRO GLU GLU TYS LEU MODRES 1C5N TYS I 63 TYR SULFONATED TYROSINE HET TYS I 63 24 HET NA 409 1 HET CA 521 1 HET ESI 246 21 HETNAM TYS SULFONATED TYROSINE HETNAM NA SODIUM ION HETNAM CA CALCIUM ION HETNAM ESI 4-IODOBENZO[B]THIOPHENE-2-CARBOXAMIDINE FORMUL 3 TYS C9 H11 N O6 S FORMUL 4 NA NA 1+ FORMUL 5 CA CA 2+ FORMUL 6 ESI C9 H8 I N2 S 1+ FORMUL 7 HOH *255(H2 O) HELIX 1 1 PHE L 7 SER L 11 5 5 HELIX 2 3 ALA H 55 CYS H 58 5 4 HELIX 3 6 ASP H 125 LEU H 130 1 9 HELIX 4 7 GLU H 164 ASP H 170 1 7 HELIX 5 9 LEU H 234 GLY H 246 1 13 SHEET 1 A 7 SER H 20 ASP H 21 0 SHEET 2 A 7 GLN H 156 PRO H 161 -1 O VAL H 157 N SER H 20 SHEET 3 A 7 LYS H 135 GLY H 140 -1 O GLY H 136 N LEU H 160 SHEET 4 A 7 PRO H 198 LYS H 202 -1 O PRO H 198 N THR H 139 SHEET 5 A 7 TRP H 207 TRP H 215 -1 N TYR H 208 O MET H 201 SHEET 6 A 7 GLY H 226 HIS H 230 -1 N PHE H 227 O TRP H 215 SHEET 7 A 7 MET H 180 ALA H 183 -1 O PHE H 181 N TYR H 228 SHEET 1 B 7 LYS H 81 SER H 83 0 SHEET 2 B 7 LEU H 64 ILE H 68 -1 O VAL H 66 N SER H 83 SHEET 3 B 7 GLN H 30 ARG H 35 -1 O MET H 32 N ARG H 67 SHEET 4 B 7 GLU H 39 LEU H 46 -1 O GLU H 39 N ARG H 35 SHEET 5 B 7 TRP H 51 THR H 54 -1 N LEU H 53 O SER H 45 SHEET 6 B 7 ALA H 104 LEU H 108 -1 O ALA H 104 N THR H 54 SHEET 7 B 7 LEU H 85 ILE H 90 -1 N GLU H 86 O LYS H 107 SHEET 1 C 2 LEU H 60 TYR H 60A 0 SHEET 2 C 2 LYS H 60F ASN H 60G-1 O LYS H 60F N TYR H 60A SSBOND 1 CYS L 1 CYS H 122 SSBOND 2 CYS H 42 CYS H 58 SSBOND 3 CYS H 168 CYS H 182 SSBOND 4 CYS H 191 CYS H 220 CISPEP 1 SER H 36A PRO H 37 0 -10.46 CRYST1 72.140 72.260 73.090 90.00 101.23 90.00 C 1 2 1 4 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 0.013862 0.000000 0.002752 0.00000 SCALE2 0.000000 0.013839 0.000000 0.00000 SCALE3 0.000000 0.000000 0.013949 0.00000