HEADER TRANSFERASE/APOPTOSIS/IMMUNE SYSTEM 12-FEB-14 4ORZ TITLE HIV-1 NEF PROTEIN IN COMPLEX WITH SINGLE DOMAIN ANTIBODY SDAB19 AND AN TITLE 2 ENGINEERED HCK SH3 DOMAIN COMPND MOL_ID: 1; COMPND 2 MOLECULE: TYROSINE-PROTEIN KINASE HCK; COMPND 3 CHAIN: A; COMPND 4 FRAGMENT: SH3 DOMAIN, UNP RESIDUES 77-138; COMPND 5 SYNONYM: HEMATOPOIETIC CELL KINASE, HEMOPOIETIC CELL KINASE, P59- COMPND 6 HCK/P60-HCK, P59HCK, P61HCK; COMPND 7 EC: 2.7.10.2; COMPND 8 ENGINEERED: YES; COMPND 9 MUTATION: YES; COMPND 10 MOL_ID: 2; COMPND 11 MOLECULE: PROTEIN NEF; COMPND 12 CHAIN: B; COMPND 13 FRAGMENT: NEF PROTEIN, UNP RESIDUES 45-210; COMPND 14 SYNONYM: 3'ORF, NEGATIVE FACTOR, F-PROTEIN, C-TERMINAL CORE PROTEIN; COMPND 15 ENGINEERED: YES; COMPND 16 MUTATION: YES; COMPND 17 MOL_ID: 3; COMPND 18 MOLECULE: SINGLE DOMAIN ANTIBODY SDAB19; COMPND 19 CHAIN: C; COMPND 20 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 3 ORGANISM_COMMON: HUMAN; SOURCE 4 ORGANISM_TAXID: 9606; SOURCE 5 GENE: HCK; SOURCE 6 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 7 EXPRESSION_SYSTEM_TAXID: 469008; SOURCE 8 EXPRESSION_SYSTEM_STRAIN: BL21(DE3); SOURCE 9 EXPRESSION_SYSTEM_VECTOR_TYPE: PGEX-2T; SOURCE 10 MOL_ID: 2; SOURCE 11 ORGANISM_SCIENTIFIC: HIV-1 M:B_ARV2/SF2; SOURCE 12 ORGANISM_COMMON: HIV-1; SOURCE 13 ORGANISM_TAXID: 11685; SOURCE 14 GENE: NEF; SOURCE 15 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 16 EXPRESSION_SYSTEM_TAXID: 469008; SOURCE 17 EXPRESSION_SYSTEM_STRAIN: BL21(DE3); SOURCE 18 EXPRESSION_SYSTEM_VECTOR_TYPE: PGEX-4T1; SOURCE 19 MOL_ID: 3; SOURCE 20 ORGANISM_SCIENTIFIC: LAMA GLAMA; SOURCE 21 ORGANISM_TAXID: 9844; SOURCE 22 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 23 EXPRESSION_SYSTEM_TAXID: 469008; SOURCE 24 EXPRESSION_SYSTEM_STRAIN: BL21(DE3); SOURCE 25 EXPRESSION_SYSTEM_VECTOR_TYPE: PGEX-4T1 KEYWDS SH3 DOMAIN, IMMUNGLOBOLIN FOLD, ANTIBODIES, EPITOPES, HIV ANTIBODIES, KEYWDS 2 HIV ACCESSORY PROTEINS, PXXP MOTIF, COMPLEMENTARITY DETERMINING KEYWDS 3 REGIONS, TRANSFERASE-APOPTOSIS-IMMUNE SYSTEM COMPLEX EXPDTA X-RAY DIFFRACTION AUTHOR M.GEYER,S.LULF REVDAT 1 26-MAR-14 4ORZ 0 JRNL AUTH S.LULF,J.MATZ,M.C.ROUYEZ,A.JARVILUOMA,K.SAKSELA,S.BENICHOU, JRNL AUTH 2 M.GEYER JRNL TITL STRUCTURAL BASIS FOR THE INHIBITION OF HIV-1 NEF BY A JRNL TITL 2 HIGH-AFFINITY BINDING SINGLE-DOMAIN ANTIBODY. JRNL REF RETROVIROLOGY V. 11 24 2014 JRNL REFN ESSN 1742-4690 JRNL PMID 24620746 JRNL DOI 10.1186/1742-4690-11-24 REMARK 2 REMARK 2 RESOLUTION. 2.00 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : PHENIX (PHENIX.REFINE: 1.8.1_1168) REMARK 3 AUTHORS : PAUL ADAMS,PAVEL AFONINE,VICENT CHEN,IAN REMARK 3 : DAVIS,KRESHNA GOPAL,RALF GROSSE- REMARK 3 : KUNSTLEVE,LI-WEI HUNG,ROBERT IMMORMINO, REMARK 3 : TOM IOERGER,AIRLIE MCCOY,ERIK MCKEE,NIGEL REMARK 3 : MORIARTY,REETAL PAI,RANDY READ,JANE REMARK 3 : RICHARDSON,DAVID RICHARDSON,TOD ROMO,JIM REMARK 3 : SACCHETTINI,NICHOLAS SAUTER,JACOB SMITH, REMARK 3 : LAURENT STORONI,TOM TERWILLIGER,PETER REMARK 3 : ZWART REMARK 3 REMARK 3 REFINEMENT TARGET : MLHL REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 2.00 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 41.75 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 2.040 REMARK 3 COMPLETENESS FOR RANGE (%) : 98.9 REMARK 3 NUMBER OF REFLECTIONS : 25031 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 R VALUE (WORKING + TEST SET) : 0.204 REMARK 3 R VALUE (WORKING SET) : 0.202 REMARK 3 FREE R VALUE : 0.236 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.000 REMARK 3 FREE R VALUE TEST SET COUNT : 1252 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 41.7600 - 4.1590 1.00 2717 144 0.1686 0.2010 REMARK 3 2 4.1590 - 3.3015 0.99 2656 139 0.1784 0.2194 REMARK 3 3 3.3015 - 2.8843 0.99 2658 140 0.2162 0.2391 REMARK 3 4 2.8843 - 2.6206 0.99 2621 138 0.2189 0.2412 REMARK 3 5 2.6206 - 2.4328 0.99 2645 139 0.2349 0.2651 REMARK 3 6 2.4328 - 2.2894 0.99 2638 139 0.2387 0.2992 REMARK 3 7 2.2894 - 2.1748 0.98 2624 138 0.2342 0.2715 REMARK 3 8 2.1748 - 2.0801 0.99 2592 137 0.2397 0.2712 REMARK 3 9 2.0801 - 2.0000 0.98 2628 138 0.2500 0.2915 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) : 0.210 REMARK 3 PHASE ERROR (DEGREES, MAXIMUM-LIKELIHOOD BASED) : 24.910 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 : 0.008 2399 REMARK 3 ANGLE : 1.288 3255 REMARK 3 CHIRALITY : 0.085 336 REMARK 3 PLANARITY : 0.006 413 REMARK 3 DIHEDRAL : 15.518 865 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 4ORZ COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 26-FEB-14. REMARK 100 THE RCSB ID CODE IS RCSB084876. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 10-FEB-12 REMARK 200 TEMPERATURE (KELVIN) : 100 REMARK 200 PH : 9.0 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : SLS REMARK 200 BEAMLINE : X10SA REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.979400 REMARK 200 MONOCHROMATOR : DIAMOND(111) REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : CCD REMARK 200 DETECTOR MANUFACTURER : PSI PILATUS 6M REMARK 200 INTENSITY-INTEGRATION SOFTWARE : XDS REMARK 200 DATA SCALING SOFTWARE : XSCALE REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 47918 REMARK 200 RESOLUTION RANGE HIGH (A) : 2.000 REMARK 200 RESOLUTION RANGE LOW (A) : 41.750 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : -3.000 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 100.0 REMARK 200 DATA REDUNDANCY : NULL REMARK 200 R MERGE (I) : NULL REMARK 200 R SYM (I) : NULL REMARK 200 FOR THE DATA SET : NULL REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.10 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.15 REMARK 200 COMPLETENESS FOR SHELL (%) : 100.0 REMARK 200 DATA REDUNDANCY IN SHELL : NULL REMARK 200 R MERGE FOR SHELL (I) : NULL REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 FOR SHELL : NULL 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: PDB ENTRY 3RBB REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 51.31 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.53 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: ABOUT 0.1 MICRO L OF PROTEIN SOLUTION REMARK 280 AT 10 MG/ML CONCENTRATION WAS MIXED WITH 0.1 MICRO L OF RESERVOIR REMARK 280 SOLUTION FROM A 70 L RESERVOIR IN 96-WELL HAMPTON 3553 REMARK 280 CRYSTALLIZATION PLATES. INITIAL CRYSTALS OF NEFSF2 SDAB19 SH3B6 REMARK 280 COULD BE OBTAINED IN 0.2 M POTASSIUM FORMATE AND 20% POLYETHYLENE REMARK 280 GLYCOL (PEG) 3350. CRYSTAL CONDITIONS WERE OPTIMIZED TO 0.2 M REMARK 280 POTASSIUM FORMATE, 17.5% POLYETHYLENE GLYCOL (PEG) 3350 AND 0.35 REMARK 280 M AMMONIUM CHLORIDE GROWN BY HANGING-DROP VAPOR DIFFUSION IN REMARK 280 LINBRO CRYSTALLIZATION PLATES. , PH 9.0, VAPOR DIFFUSION, HANGING REMARK 280 DROP, TEMPERATURE 293K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 41 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -X,-Y,Z+1/2 REMARK 290 3555 -Y,X,Z+1/4 REMARK 290 4555 Y,-X,Z+3/4 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 35.50000 REMARK 290 SMTRY1 3 0.000000 -1.000000 0.000000 0.00000 REMARK 290 SMTRY2 3 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 17.75000 REMARK 290 SMTRY1 4 0.000000 1.000000 0.000000 0.00000 REMARK 290 SMTRY2 4 -1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY3 4 0.000000 0.000000 1.000000 53.25000 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: TRIMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, B, C 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 GLY A 72 REMARK 465 ALA A 73 REMARK 465 HIS A 74 REMARK 465 MET A 75 REMARK 465 GLY A 76 REMARK 465 GLY A 77 REMARK 465 SER A 78 REMARK 465 GLU A 79 REMARK 465 ASP A 80 REMARK 465 ASP A 137 REMARK 465 SER A 138 REMARK 465 GLY B 45 REMARK 465 ALA B 46 REMARK 465 MET B 47 REMARK 465 ALA B 48 REMARK 465 SER B 49 REMARK 465 SER B 50 REMARK 465 ASN B 51 REMARK 465 THR B 52 REMARK 465 ALA B 53 REMARK 465 ALA B 54 REMARK 465 THR B 55 REMARK 465 ASN B 56 REMARK 465 ALA B 57 REMARK 465 ASP B 58 REMARK 465 SER B 59 REMARK 465 ALA B 60 REMARK 465 TRP B 61 REMARK 465 LEU B 62 REMARK 465 GLU B 63 REMARK 465 ALA B 64 REMARK 465 GLN B 65 REMARK 465 GLU B 66 REMARK 465 GLU B 67 REMARK 465 GLU B 68 REMARK 465 GLU B 69 REMARK 465 VAL B 70 REMARK 465 ASP B 209 REMARK 465 ALA B 210 REMARK 465 GLY C -1 REMARK 465 ALA C 0 REMARK 465 MET C 1 REMARK 465 ALA C 2 REMARK 465 GLU C 3 REMARK 465 VAL C 4 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 DISTANCE REMARK 500 O HOH B 328 O HOH B 381 1.91 REMARK 500 O HOH B 324 O HOH B 327 1.99 REMARK 500 O HOH B 349 O HOH B 379 2.07 REMARK 500 O HOH C 204 O HOH C 207 2.11 REMARK 500 NH1 ARG C 99 OD2 ASP C 106 2.12 REMARK 500 O HOH B 371 O HOH B 373 2.16 REMARK 500 OD1 ASP C 63 O HOH B 390 2.17 REMARK 500 NH1 ARG B 109 OE2 GLU B 112 2.18 REMARK 500 REMARK 500 REMARK: NULL 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 GLY C 28 N - CA - C ANGL. DEV. = -15.6 DEGREES REMARK 500 PHE C 29 N - CA - C ANGL. DEV. = -19.2 DEGREES REMARK 500 LEU C 39 CA - CB - CG ANGL. DEV. = 14.3 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 A 109 113.93 -163.62 REMARK 500 SER A 111 38.12 -89.01 REMARK 500 PHE C 29 129.14 157.73 REMARK 500 SER C 56 34.51 -97.02 REMARK 500 SER C 64 1.93 -58.31 REMARK 500 LYS C 66 -121.39 46.24 REMARK 500 ALA C 93 172.14 178.25 REMARK 500 REMARK 500 REMARK: NULL REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: 3RBB RELATED DB: PDB REMARK 900 HIV-1 NEF PROTEIN IN COMPLEX WITH ENGINEERED HCK SH3 DOMAIN REMARK 900 RELATED ID: 3REA RELATED DB: PDB REMARK 900 HIV-1 NEF PROTEIN IN COMPLEX WITH ENGINEERED HCK-SH3 DOMAIN DBREF 4ORZ A 77 138 UNP P08631 HCK_HUMAN 77 138 DBREF 4ORZ B 45 210 UNP P03407 NEF_HV1A2 45 210 DBREF 4ORZ C -1 118 PDB 4ORZ 4ORZ -1 118 SEQADV 4ORZ GLY A 72 UNP P08631 EXPRESSION TAG SEQADV 4ORZ ALA A 73 UNP P08631 EXPRESSION TAG SEQADV 4ORZ HIS A 74 UNP P08631 EXPRESSION TAG SEQADV 4ORZ MET A 75 UNP P08631 EXPRESSION TAG SEQADV 4ORZ GLY A 76 UNP P08631 EXPRESSION TAG SEQADV 4ORZ TYR A 90 UNP P08631 GLU 90 ENGINEERED MUTATION SEQADV 4ORZ SER A 91 UNP P08631 ALA 91 ENGINEERED MUTATION SEQADV 4ORZ PRO A 92 UNP P08631 ILE 92 ENGINEERED MUTATION SEQADV 4ORZ PHE A 93 UNP P08631 HIS 93 ENGINEERED MUTATION SEQADV 4ORZ SER A 94 UNP P08631 HIS 94 ENGINEERED MUTATION SEQADV 4ORZ TRP A 95 UNP P08631 GLU 95 ENGINEERED MUTATION SEQADV 4ORZ MET B 47 UNP P03407 ILE 47 ENGINEERED MUTATION SEQADV 4ORZ ALA B 48 UNP P03407 THR 48 ENGINEERED MUTATION SEQADV 4ORZ SER B 59 UNP P03407 CYS 59 ENGINEERED MUTATION SEQADV 4ORZ B UNP P03407 GLU 158 DELETION SEQADV 4ORZ B UNP P03407 GLU 159 DELETION SEQADV 4ORZ B UNP P03407 ALA 160 DELETION SEQADV 4ORZ B UNP P03407 ASN 161 DELETION SEQADV 4ORZ B UNP P03407 GLU 162 DELETION SEQADV 4ORZ B UNP P03407 GLY 163 DELETION SEQADV 4ORZ B UNP P03407 GLU 164 DELETION SEQADV 4ORZ B UNP P03407 ASN 165 DELETION SEQADV 4ORZ B UNP P03407 ASN 166 DELETION SEQADV 4ORZ B UNP P03407 SER 167 DELETION SEQADV 4ORZ B UNP P03407 LEU 168 DELETION SEQADV 4ORZ B UNP P03407 LEU 169 DELETION SEQADV 4ORZ B UNP P03407 HIS 170 DELETION SEQADV 4ORZ B UNP P03407 PRO 171 DELETION SEQADV 4ORZ B UNP P03407 MET 172 DELETION SEQADV 4ORZ B UNP P03407 SER 173 DELETION SEQADV 4ORZ B UNP P03407 LEU 174 DELETION SEQADV 4ORZ B UNP P03407 HIS 175 DELETION SEQADV 4ORZ B UNP P03407 GLY 176 DELETION SEQADV 4ORZ B UNP P03407 MET 177 DELETION SEQADV 4ORZ B UNP P03407 GLU 178 DELETION SEQADV 4ORZ ALA B 210 UNP P03407 CYS 210 ENGINEERED MUTATION SEQRES 1 A 67 GLY ALA HIS MET GLY GLY SER GLU ASP ILE ILE VAL VAL SEQRES 2 A 67 ALA LEU TYR ASP TYR TYR SER PRO PHE SER TRP ASP LEU SEQRES 3 A 67 SER PHE GLN LYS GLY ASP GLN MET VAL VAL LEU GLU GLU SEQRES 4 A 67 SER GLY GLU TRP TRP LYS ALA ARG SER LEU ALA THR ARG SEQRES 5 A 67 LYS GLU GLY TYR ILE PRO SER ASN TYR VAL ALA ARG VAL SEQRES 6 A 67 ASP SER SEQRES 1 B 145 GLY ALA MET ALA SER SER ASN THR ALA ALA THR ASN ALA SEQRES 2 B 145 ASP SER ALA TRP LEU GLU ALA GLN GLU GLU GLU GLU VAL SEQRES 3 B 145 GLY PHE PRO VAL ARG PRO GLN VAL PRO LEU ARG PRO MET SEQRES 4 B 145 THR TYR LYS ALA ALA LEU ASP ILE SER HIS PHE LEU LYS SEQRES 5 B 145 GLU LYS GLY GLY LEU GLU GLY LEU ILE TRP SER GLN ARG SEQRES 6 B 145 ARG GLN GLU ILE LEU ASP LEU TRP ILE TYR HIS THR GLN SEQRES 7 B 145 GLY TYR PHE PRO ASP TRP GLN ASN TYR THR PRO GLY PRO SEQRES 8 B 145 GLY ILE ARG TYR PRO LEU THR PHE GLY TRP CYS PHE LYS SEQRES 9 B 145 LEU VAL PRO VAL GLU PRO GLU LYS VAL ASP ALA GLU LYS SEQRES 10 B 145 GLU VAL LEU VAL TRP ARG PHE ASP SER LYS LEU ALA PHE SEQRES 11 B 145 HIS HIS MET ALA ARG GLU LEU HIS PRO GLU TYR TYR LYS SEQRES 12 B 145 ASP ALA SEQRES 1 C 120 GLY ALA MET ALA GLU VAL GLN LEU VAL GLU SER GLY GLY SEQRES 2 C 120 GLY LEU VAL GLN ALA GLY GLY SER LEU ARG LEU PHE CYS SEQRES 3 C 120 ALA ALA SER GLY PHE THR PHE GLY THR SER ASN MET ALA SEQRES 4 C 120 TRP LEU ARG GLN ALA PRO GLY LYS ARG ARG GLU TRP VAL SEQRES 5 C 120 ALA LEU ILE THR ILE SER GLY TYR THR ASP TYR ALA ASP SEQRES 6 C 120 SER VAL LYS ASP ARG PHE THR ILE SER ARG ASP ASN ALA SEQRES 7 C 120 LYS ASN THR VAL SER LEU GLN MET ASN SER LEU LYS PRO SEQRES 8 C 120 GLU ASP THR ALA ILE TYR PHE CYS ALA ARG ARG VAL GLY SEQRES 9 C 120 SER GLU TYR ASP LEU TRP GLY GLN GLY THR GLN VAL THR SEQRES 10 C 120 VAL SER SER FORMUL 4 HOH *125(H2 O) HELIX 1 1 THR B 84 GLY B 99 1 16 HELIX 2 2 SER B 107 GLY B 123 1 17 HELIX 3 3 ASP B 179 GLU B 183 5 5 HELIX 4 4 SER B 191 PHE B 195 5 5 HELIX 5 5 HIS B 197 HIS B 203 1 7 HELIX 6 6 PRO B 204 TYR B 207 5 4 HELIX 7 7 ASP C 63 LYS C 66 5 4 HELIX 8 8 LYS C 88 THR C 92 5 5 SHEET 1 A 5 GLU A 125 PRO A 129 0 SHEET 2 A 5 TRP A 114 SER A 119 -1 N ALA A 117 O GLY A 126 SHEET 3 A 5 GLN A 104 GLU A 109 -1 N VAL A 106 O ARG A 118 SHEET 4 A 5 ILE A 82 ALA A 85 -1 N VAL A 83 O MET A 105 SHEET 5 A 5 VAL A 133 ARG A 135 -1 O ALA A 134 N VAL A 84 SHEET 1 B 2 PHE B 147 PRO B 151 0 SHEET 2 B 2 LEU B 185 PHE B 189 -1 O VAL B 186 N VAL B 150 SHEET 1 C 4 LEU C 6 SER C 9 0 SHEET 2 C 4 LEU C 20 ALA C 26 -1 O PHE C 23 N SER C 9 SHEET 3 C 4 THR C 79 MET C 84 -1 O LEU C 82 N LEU C 22 SHEET 4 C 4 PHE C 69 ASP C 74 -1 N THR C 70 O GLN C 83 SHEET 1 D 6 GLY C 12 GLN C 15 0 SHEET 2 D 6 THR C 112 SER C 117 1 O THR C 115 N VAL C 14 SHEET 3 D 6 ALA C 93 VAL C 101 -1 N TYR C 95 O THR C 112 SHEET 4 D 6 ASN C 35 GLN C 41 -1 N LEU C 39 O PHE C 96 SHEET 5 D 6 GLU C 48 ILE C 53 -1 O ILE C 53 N MET C 36 SHEET 6 D 6 THR C 59 TYR C 61 -1 O ASP C 60 N LEU C 52 SHEET 1 E 4 GLY C 12 GLN C 15 0 SHEET 2 E 4 THR C 112 SER C 117 1 O THR C 115 N VAL C 14 SHEET 3 E 4 ALA C 93 VAL C 101 -1 N TYR C 95 O THR C 112 SHEET 4 E 4 GLU C 104 TRP C 108 -1 O GLU C 104 N VAL C 101 CISPEP 1 GLY B 134 PRO B 135 0 10.96 CISPEP 2 PRO B 154 GLU B 155 0 0.81 CISPEP 3 GLY C 28 PHE C 29 0 3.38 CRYST1 73.000 73.000 71.000 90.00 90.00 90.00 P 41 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 0.013699 0.000000 0.000000 0.00000 SCALE2 0.000000 0.013699 0.000000 0.00000 SCALE3 0.000000 0.000000 0.014085 0.00000