HEADER IMMUNE SYSTEM 09-OCT-23 8UHH TITLE ANTI-PHOSPHOHISTIDINE FAB HSC44.CK.ELBOW.20.N32F COMPND MOL_ID: 1; COMPND 2 MOLECULE: HSC44.CK.ELBOW.20.N32F FAB HEAVY CHAIN; COMPND 3 CHAIN: H; COMPND 4 ENGINEERED: YES; COMPND 5 OTHER_DETAILS: FAB HEAVY CHAIN; COMPND 6 MOL_ID: 2; COMPND 7 MOLECULE: HSC44.CK.ELBOW.20.N32F FAB LIGHT CHAIN; COMPND 8 CHAIN: L; COMPND 9 ENGINEERED: YES; COMPND 10 OTHER_DETAILS: FAB LIGHT CHAIN SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: ORYCTOLAGUS CUNICULUS; SOURCE 3 ORGANISM_TAXID: 9986; SOURCE 4 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 5 EXPRESSION_SYSTEM_TAXID: 9606; SOURCE 6 MOL_ID: 2; SOURCE 7 ORGANISM_SCIENTIFIC: ORYCTOLAGUS CUNICULUS; SOURCE 8 ORGANISM_TAXID: 9986; SOURCE 9 EXPRESSION_SYSTEM: HOMO SAPIENS; SOURCE 10 EXPRESSION_SYSTEM_TAXID: 9606 KEYWDS ANTI-PHOSPHOHISTIDINE ANTIBODY POST-TRANSLATIONAL MODIFICATION KEYWDS 2 ANTIBODY HUMANIZATION, IMMUNE SYSTEM EXPDTA X-RAY DIFFRACTION AUTHOR R.KALAGIRI,R.L.STANFIELD,T.HUNTER,I.A.WILSON REVDAT 1 19-MAR-25 8UHH 0 JRNL AUTH G.D.MARTYN,R.KALAGIRI,G.VEGGIANI,R.L.STANFIELD,I.CHOUDHURI, JRNL AUTH 2 M.SALA,J.MEISENHELDER,C.CHEN,A.BISWAS,R.M.LEVY,D.LYUMKIS, JRNL AUTH 3 I.A.WILSON,T.HUNTER,S.S.SIDHU JRNL TITL USING PHAGE DISPLAY FOR RATIONAL ENGINEERING OF A HIGHER JRNL TITL 2 AFFINITY HUMANIZED 3' PHOSPHOHISTIDINE-SPECIFIC ANTIBODY. JRNL REF BIORXIV 2024 JRNL REFN ISSN 2692-8205 JRNL PMID 39574610 JRNL DOI 10.1101/2024.11.04.621849 REMARK 2 REMARK 2 RESOLUTION. 1.84 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : PHENIX 1.19.2_4158 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 : GEOSTD + MONOMER LIBRARY + CDL V1.2 REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.84 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 46.27 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 1.340 REMARK 3 COMPLETENESS FOR RANGE (%) : 98.8 REMARK 3 NUMBER OF REFLECTIONS : 46523 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.206 REMARK 3 R VALUE (WORKING SET) : 0.204 REMARK 3 FREE R VALUE : 0.236 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.940 REMARK 3 FREE R VALUE TEST SET COUNT : 2299 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 46.2700 - 4.6400 1.00 3034 138 0.1966 0.2090 REMARK 3 2 4.6400 - 3.6800 1.00 2860 162 0.1543 0.1628 REMARK 3 3 3.6800 - 3.2200 1.00 2817 158 0.1837 0.2552 REMARK 3 4 3.2200 - 2.9200 1.00 2826 137 0.1895 0.2258 REMARK 3 5 2.9200 - 2.7200 1.00 2790 142 0.2040 0.2387 REMARK 3 6 2.7100 - 2.5500 1.00 2803 138 0.2042 0.2350 REMARK 3 7 2.5500 - 2.4300 1.00 2769 147 0.2067 0.2683 REMARK 3 8 2.4300 - 2.3200 1.00 2791 145 0.2128 0.2436 REMARK 3 9 2.3200 - 2.2300 1.00 2756 163 0.2226 0.2917 REMARK 3 10 2.2300 - 2.1500 1.00 2781 141 0.2242 0.2852 REMARK 3 11 2.1500 - 2.0900 1.00 2758 146 0.2338 0.2583 REMARK 3 12 2.0900 - 2.0300 1.00 2731 128 0.2512 0.2786 REMARK 3 13 2.0300 - 1.9700 0.99 2747 149 0.2677 0.3215 REMARK 3 14 1.9700 - 1.9300 0.98 2695 141 0.2783 0.3358 REMARK 3 15 1.9300 - 1.8800 0.96 2629 136 0.3033 0.2734 REMARK 3 16 1.8800 - 1.8400 0.88 2437 128 0.3768 0.3499 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) : 0.237 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 25.119 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 25.39 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 30.74 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 : 0.005 3436 REMARK 3 ANGLE : 0.861 4677 REMARK 3 CHIRALITY : 0.050 526 REMARK 3 PLANARITY : 0.007 594 REMARK 3 DIHEDRAL : 13.914 1238 REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : NULL 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 8UHH COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 17-OCT-23. REMARK 100 THE DEPOSITION ID IS D_1000277453. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 24-SEP-22 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 7.5 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : ALS REMARK 200 BEAMLINE : 5.0.1 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.97741 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : PIXEL REMARK 200 DETECTOR MANUFACTURER : DECTRIS PILATUS3 2M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : HKL-2000 REMARK 200 DATA SCALING SOFTWARE : HKL-2000 REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 46607 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.840 REMARK 200 RESOLUTION RANGE LOW (A) : 46.270 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : -3.000 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 99.2 REMARK 200 DATA REDUNDANCY : 6.300 REMARK 200 R MERGE (I) : 0.10000 REMARK 200 R SYM (I) : 0.10000 REMARK 200 FOR THE DATA SET : 21.0000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.84 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.88 REMARK 200 COMPLETENESS FOR SHELL (%) : 94.4 REMARK 200 DATA REDUNDANCY IN SHELL : 4.50 REMARK 200 R MERGE FOR SHELL (I) : 1.78000 REMARK 200 R SYM FOR SHELL (I) : 1.78000 REMARK 200 FOR SHELL : 0.860 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: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 56.06 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.80 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 0.1M HEPES, PH 7.5, 20% PEG4000, 10% 2 REMARK 280 -PROPANOL, VAPOR DIFFUSION, SITTING DROP, TEMPERATURE 293K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 21 21 21 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X+1/2,-Y,Z+1/2 REMARK 290 3555 -X,Y+1/2,-Z+1/2 REMARK 290 4555 X+1/2,-Y+1/2,-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 24.28700 REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 76.09450 REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 36.03600 REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 76.09450 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 24.28700 REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 36.03600 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 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: 4780 ANGSTROM**2 REMARK 350 SURFACE AREA OF THE COMPLEX: 19800 ANGSTROM**2 REMARK 350 CHANGE IN SOLVENT FREE ENERGY: -32.0 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 ASP H 216 REMARK 465 LYS H 217 REMARK 465 THR H 218 REMARK 465 HIS H 219 REMARK 465 THR H 220 REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: COVALENT BOND ANGLES REMARK 500 REMARK 500 THE STEREOCHEMICAL PARAMETERS OF THE FOLLOWING RESIDUES REMARK 500 HAVE VALUES WHICH DEVIATE FROM EXPECTED VALUES BY MORE REMARK 500 THAN 6*RMSD (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN REMARK 500 IDENTIFIER; SSEQ=SEQUENCE NUMBER; I=INSERTION CODE). REMARK 500 REMARK 500 STANDARD TABLE: REMARK 500 FORMAT: (10X,I3,1X,A3,1X,A1,I4,A1,3(1X,A4,2X),12X,F5.1) REMARK 500 REMARK 500 EXPECTED VALUES PROTEIN: ENGH AND HUBER, 1999 REMARK 500 EXPECTED VALUES NUCLEIC ACID: CLOWNEY ET AL 1996 REMARK 500 REMARK 500 M RES CSSEQI ATM1 ATM2 ATM3 REMARK 500 CYS L 194 CA - CB - SG ANGL. DEV. = 8.7 DEGREES REMARK 500 REMARK 500 REMARK: NULL 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 GLU H 85 -9.89 -59.56 REMARK 500 ASN L 30 15.04 57.33 REMARK 500 ILE L 51 -54.37 72.82 REMARK 500 SER L 67 144.80 -170.65 REMARK 500 ASN L 138 72.23 51.86 REMARK 500 REMARK 500 REMARK: NULL REMARK 525 REMARK 525 SOLVENT REMARK 525 REMARK 525 THE SOLVENT MOLECULES HAVE CHAIN IDENTIFIERS THAT REMARK 525 INDICATE THE POLYMER CHAIN WITH WHICH THEY ARE MOST REMARK 525 CLOSELY ASSOCIATED. THE REMARK LISTS ALL THE SOLVENT REMARK 525 MOLECULES WHICH ARE MORE THAN 5A AWAY FROM THE REMARK 525 NEAREST POLYMER CHAIN (M = MODEL NUMBER; REMARK 525 RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE REMARK 525 NUMBER; I=INSERTION CODE): REMARK 525 REMARK 525 M RES CSSEQI REMARK 525 HOH H 609 DISTANCE = 5.81 ANGSTROMS REMARK 525 HOH H 610 DISTANCE = 6.61 ANGSTROMS REMARK 525 HOH L 648 DISTANCE = 6.32 ANGSTROMS REMARK 525 HOH L 649 DISTANCE = 7.24 ANGSTROMS DBREF 8UHH H 1 220 PDB 8UHH 8UHH 1 220 DBREF 8UHH L 1 212 PDB 8UHH 8UHH 1 212 SEQRES 1 H 224 GLU VAL GLN LEU VAL GLU SER GLY GLY GLY LEU VAL GLN SEQRES 2 H 224 PRO GLY GLY SER LEU ARG LEU SER CYS THR ALA SER GLY SEQRES 3 H 224 PHE SER ILE ASP SER TYR GLY PHE SER TRP VAL ARG GLN SEQRES 4 H 224 ALA PRO GLY LYS GLY LEU GLU HIS ILE GLY TYR LEU THR SEQRES 5 H 224 ALA GLY GLY ARG ALA PHE TYR ALA SER TRP ALA LYS SER SEQRES 6 H 224 ARG SER THR ILE THR ARG ASN THR ASN GLU ASN THR VAL SEQRES 7 H 224 THR LEU LYS MET ASN SER LEU ARG ALA GLU ASP THR ALA SEQRES 8 H 224 VAL TYR TYR CYS ALA LYS LEU GLY THR GLY SER ARG PHE SEQRES 9 H 224 ALA ILE TRP GLY GLN GLY THR LEU VAL THR VAL PHE ASN SEQRES 10 H 224 GLN ILE LYS GLY PRO SER VAL PHE PRO LEU ALA PRO SER SEQRES 11 H 224 SER LYS SER THR SER GLY GLY THR ALA ALA LEU GLY CYS SEQRES 12 H 224 LEU VAL LYS ASP TYR PHE PRO GLU PRO VAL THR VAL SER SEQRES 13 H 224 TRP ASN SER GLY ALA LEU THR SER GLY VAL HIS THR PHE SEQRES 14 H 224 PRO ALA VAL LEU GLN SER SER GLY LEU TYR SER LEU SER SEQRES 15 H 224 SER VAL VAL THR VAL PRO SER SER SER LEU GLY THR GLN SEQRES 16 H 224 THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SER ASN THR SEQRES 17 H 224 LYS VAL ASP LYS LYS VAL GLU PRO LYS SER CYS ASP LYS SEQRES 18 H 224 THR HIS THR SEQRES 1 L 218 ASP ILE GLN MET THR GLN SER PRO SER SER LEU SER ALA SEQRES 2 L 218 SER VAL GLY ASP ARG VAL THR ILE THR CYS ARG ALA SER SEQRES 3 L 218 GLN SER VAL TRP ARG ASN LYS PHE VAL ALA TRP TYR GLN SEQRES 4 L 218 GLN LYS PRO GLY LYS ALA PRO LYS ARG LEU ILE TYR ALA SEQRES 5 L 218 ILE ALA SER LEU TYR SER GLY VAL PRO SER ARG PHE SER SEQRES 6 L 218 GLY SER GLY SER GLY THR ASP PHE THR LEU THR ILE SER SEQRES 7 L 218 SER LEU GLN PRO GLU ASP PHE ALA THR TYR TYR CYS VAL SEQRES 8 L 218 GLY HIS TYR GLY SER GLU ASN ASP ALA TYR TYR ALA PHE SEQRES 9 L 218 GLY GLN GLY THR LYS VAL GLU ILE LYS ARG THR VAL ALA SEQRES 10 L 218 ALA PRO SER VAL PHE ILE PHE PRO PRO SER ASP GLU GLN SEQRES 11 L 218 LEU LYS SER GLY THR ALA SER VAL VAL CYS LEU LEU ASN SEQRES 12 L 218 ASN PHE TYR PRO ARG GLU ALA LYS VAL SER TRP TYR VAL SEQRES 13 L 218 ASP ASN ALA LEU GLN SER GLY ASN SER GLN GLU SER VAL SEQRES 14 L 218 THR GLU GLN ASP SER LYS ASP SER THR TYR SER LEU SER SEQRES 15 L 218 SER THR LEU THR LEU SER LYS ALA ASP TYR GLU LYS HIS SEQRES 16 L 218 LYS VAL TYR ALA CYS GLU VAL THR GLN GLY THR THR SER SEQRES 17 L 218 VAL THR LYS SER PHE ASN ARG GLY GLU CYS HET EPE H 301 32 HET EDO H 302 10 HET CL H 303 1 HET CL L 301 1 HETNAM EPE 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID HETNAM EDO 1,2-ETHANEDIOL HETNAM CL CHLORIDE ION HETSYN EPE HEPES HETSYN EDO ETHYLENE GLYCOL FORMUL 3 EPE C8 H18 N2 O4 S FORMUL 4 EDO C2 H6 O2 FORMUL 5 CL 2(CL 1-) FORMUL 7 HOH *459(H2 O) HELIX 1 AA1 SER H 28 TYR H 32 5 5 HELIX 2 AA2 ALA H 60 SER H 65 1 6 HELIX 3 AA3 ARG H 83 THR H 87 5 5 HELIX 4 AA4 SER H 155 ALA H 157 5 3 HELIX 5 AA5 SER H 186 LEU H 188 5 3 HELIX 6 AA6 LYS H 200 ASN H 203 5 4 HELIX 7 AA7 VAL L 27B LYS L 31 5 5 HELIX 8 AA8 GLN L 79 PHE L 83 5 5 HELIX 9 AA9 SER L 121 LYS L 126 1 6 HELIX 10 AB1 LYS L 183 LYS L 188 1 6 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 VAL H 5 SHEET 3 AA1 4 THR H 77 MET H 82 -1 O MET H 82 N LEU H 18 SHEET 4 AA1 4 SER H 67 ASN H 72 -1 N THR H 70 O THR H 79 SHEET 1 AA2 6 GLY H 10 VAL H 12 0 SHEET 2 AA2 6 THR H 107 VAL H 111 1 O THR H 110 N VAL H 12 SHEET 3 AA2 6 ALA H 88 LYS H 94 -1 N TYR H 90 O THR H 107 SHEET 4 AA2 6 PHE H 34 GLN H 39 -1 N SER H 35 O ALA H 93 SHEET 5 AA2 6 LEU H 45 LEU H 51 -1 O ILE H 48 N TRP H 36 SHEET 6 AA2 6 ALA H 57 TYR H 59 -1 O PHE H 58 N TYR H 50 SHEET 1 AA3 4 GLY H 10 VAL H 12 0 SHEET 2 AA3 4 THR H 107 VAL H 111 1 O THR H 110 N VAL H 12 SHEET 3 AA3 4 ALA H 88 LYS H 94 -1 N TYR H 90 O THR H 107 SHEET 4 AA3 4 ILE H 102 TRP H 103 -1 O ILE H 102 N LYS H 94 SHEET 1 AA4 4 SER H 119 LEU H 123 0 SHEET 2 AA4 4 THR H 134 TYR H 144 -1 O GLY H 138 N LEU H 123 SHEET 3 AA4 4 TYR H 175 PRO H 184 -1 O LEU H 177 N VAL H 141 SHEET 4 AA4 4 VAL H 162 THR H 164 -1 N HIS H 163 O VAL H 180 SHEET 1 AA5 4 SER H 119 LEU H 123 0 SHEET 2 AA5 4 THR H 134 TYR H 144 -1 O GLY H 138 N LEU H 123 SHEET 3 AA5 4 TYR H 175 PRO H 184 -1 O LEU H 177 N VAL H 141 SHEET 4 AA5 4 VAL H 168 LEU H 169 -1 N VAL H 168 O SER H 176 SHEET 1 AA6 3 THR H 150 TRP H 153 0 SHEET 2 AA6 3 ILE H 194 HIS H 199 -1 O ASN H 196 N SER H 152 SHEET 3 AA6 3 THR H 204 LYS H 209 -1 O VAL H 206 N VAL H 197 SHEET 1 AA7 4 MET L 4 SER L 7 0 SHEET 2 AA7 4 VAL L 19 ALA L 25 -1 O ARG L 24 N THR L 5 SHEET 3 AA7 4 ASP L 70 ILE L 75 -1 O LEU 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 SER L 94 -1 N ALA L 84 O VAL L 104 SHEET 4 AA8 6 VAL L 33 GLN L 38 -1 N TYR L 36 O TYR L 87 SHEET 5 AA8 6 LYS L 45 TYR L 49 -1 O LYS L 45 N GLN L 37 SHEET 6 AA8 6 SER L 53 LEU L 54 -1 O SER 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 SER L 94 -1 N ALA L 84 O VAL L 104 SHEET 4 AA9 4 ASP L 95B PHE L 98 -1 O TYR L 95D N TYR L 92 SHEET 1 AB1 4 SER L 114 PHE L 118 0 SHEET 2 AB1 4 THR L 129 PHE L 139 -1 O ASN L 137 N SER L 114 SHEET 3 AB1 4 TYR L 173 SER L 182 -1 O LEU L 181 N ALA L 130 SHEET 4 AB1 4 SER L 159 VAL L 163 -1 N SER L 162 O SER L 176 SHEET 1 AB2 4 ALA L 153 LEU L 154 0 SHEET 2 AB2 4 LYS L 145 VAL L 150 -1 N VAL L 150 O ALA L 153 SHEET 3 AB2 4 VAL L 191 GLN L 198 -1 O THR L 197 N LYS L 145 SHEET 4 AB2 4 THR L 201 ASN L 208 -1 O VAL L 203 N VAL L 196 SSBOND 1 CYS H 22 CYS H 92 1555 1555 2.04 SSBOND 2 CYS H 139 CYS H 195 1555 1555 2.03 SSBOND 3 CYS H 215 CYS L 212 1555 1555 2.04 SSBOND 4 CYS L 23 CYS L 88 1555 1555 2.06 SSBOND 5 CYS L 134 CYS L 194 1555 1555 2.00 CISPEP 1 PHE H 145 PRO H 146 0 -6.28 CISPEP 2 GLU H 147 PRO H 148 0 -1.07 CISPEP 3 SER L 7 PRO L 8 0 -6.38 CISPEP 4 TYR L 140 PRO L 141 0 0.97 CRYST1 48.574 72.072 152.189 90.00 90.00 90.00 P 21 21 21 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 0.020587 0.000000 0.000000 0.00000 SCALE2 0.000000 0.013875 0.000000 0.00000 SCALE3 0.000000 0.000000 0.006571 0.00000