HEADER MEMBRANE PROTEIN/IMMUNE SYSYTEM 15-APR-24 9BEI TITLE CRYO-EM STRUCTURE OF SYNTHETIC CLAUDIN-4 COMPLEX WITH CLOSTRIDIUM TITLE 2 PERFRINGENS ENTEROTOXIN C-TERMINAL DOMAIN, SFAB COP-2, AND NANOBODY COMPND MOL_ID: 1; COMPND 2 MOLECULE: CLAUDIN-4; COMPND 3 CHAIN: A; COMPND 4 ENGINEERED: YES; COMPND 5 MOL_ID: 2; COMPND 6 MOLECULE: HEAT-LABILE ENTEROTOXIN B CHAIN; COMPND 7 CHAIN: B; COMPND 8 ENGINEERED: YES; COMPND 9 MOL_ID: 3; COMPND 10 MOLECULE: COP-2 FAB HEAVY CHAIN; COMPND 11 CHAIN: H; COMPND 12 ENGINEERED: YES; COMPND 13 MOL_ID: 4; COMPND 14 MOLECULE: ANTI-FAB NANOBODY; COMPND 15 CHAIN: K; COMPND 16 ENGINEERED: YES; COMPND 17 MOL_ID: 5; COMPND 18 MOLECULE: COP-2 FAB LIGHT CHAIN; COMPND 19 CHAIN: L; COMPND 20 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 SYNTHETIC: YES; SOURCE 3 ORGANISM_SCIENTIFIC: HOMO SAPIENS; SOURCE 4 ORGANISM_COMMON: HUMAN; SOURCE 5 ORGANISM_TAXID: 9606; SOURCE 6 MOL_ID: 2; SOURCE 7 ORGANISM_SCIENTIFIC: CLOSTRIDIUM PERFRINGENS; SOURCE 8 ORGANISM_TAXID: 1502; SOURCE 9 GENE: CPE; SOURCE 10 EXPRESSION_SYSTEM: TRICHOPLUSIA NI; SOURCE 11 EXPRESSION_SYSTEM_COMMON: CABBAGE LOOPER; SOURCE 12 EXPRESSION_SYSTEM_TAXID: 7111; SOURCE 13 MOL_ID: 3; SOURCE 14 SYNTHETIC: YES; SOURCE 15 ORGANISM_SCIENTIFIC: ESCHERICHIA COLI; SOURCE 16 ORGANISM_TAXID: 562; SOURCE 17 MOL_ID: 4; SOURCE 18 SYNTHETIC: YES; SOURCE 19 ORGANISM_SCIENTIFIC: ESCHERICHIA COLI; SOURCE 20 ORGANISM_TAXID: 562; SOURCE 21 MOL_ID: 5; SOURCE 22 SYNTHETIC: YES; SOURCE 23 ORGANISM_SCIENTIFIC: ESCHERICHIA COLI; SOURCE 24 ORGANISM_TAXID: 562 KEYWDS CLAUDIN, FAB, TOXIN, MEMBRANE PROTEIN, MEMBRANE PROTEIN-IMMUNE KEYWDS 2 SYSYTEM COMPLEX EXPDTA ELECTRON MICROSCOPY AUTHOR A.J.VECCHIO JRNL AUTH C.A.GOVERDE,M.PACESA,N.GOLDBACH,L.J.DORNFELD,P.E.BALBI, JRNL AUTH 2 S.GEORGEON,S.ROSSET,S.KAPOOR,J.CHOUDHURY,J.DEUPAREA, JRNL AUTH 3 C.SCHELLHAAS,S.KOZLOV,D.BAKER,S.OVCHINNIKOV,A.J.VECCHIO, JRNL AUTH 4 B.E.CORREIA JRNL TITL COMPUTATIONAL DESIGN OF SOLUBLE FUNCTIONAL ANALOGUES OF JRNL TITL 2 INTEGRAL MEMBRANE PROTEINS JRNL REF TO BE PUBLISHED JRNL REFN REMARK 2 REMARK 2 RESOLUTION. 4.16 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 SOFTWARE PACKAGES : CRYOSPARC, EPU, CRYOSPARC, PHENIX, REMARK 3 PHENIX, CRYOSPARC, CRYOSPARC, CRYOSPARC, REMARK 3 CRYOSPARC REMARK 3 RECONSTRUCTION SCHEMA : NULL REMARK 3 REMARK 3 EM MAP-MODEL FITTING AND REFINEMENT REMARK 3 PDB ENTRY : 7DTM REMARK 3 REFINEMENT SPACE : REAL REMARK 3 REFINEMENT PROTOCOL : FLEXIBLE FIT REMARK 3 REFINEMENT TARGET : NULL REMARK 3 OVERALL ANISOTROPIC B VALUE : 322.000 REMARK 3 REMARK 3 FITTING PROCEDURE : NULL REMARK 3 REMARK 3 EM IMAGE RECONSTRUCTION STATISTICS REMARK 3 NOMINAL PIXEL SIZE (ANGSTROMS) : NULL REMARK 3 ACTUAL PIXEL SIZE (ANGSTROMS) : NULL REMARK 3 EFFECTIVE RESOLUTION (ANGSTROMS) : 4.160 REMARK 3 NUMBER OF PARTICLES : 21296 REMARK 3 CTF CORRECTION METHOD : PHASE FLIPPING AND AMPLITUDE REMARK 3 CORRECTION REMARK 3 REMARK 3 EM RECONSTRUCTION MAGNIFICATION CALIBRATION: NULL REMARK 3 REMARK 3 OTHER DETAILS: NULL REMARK 4 REMARK 4 9BEI 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_1000283191. REMARK 245 REMARK 245 EXPERIMENTAL DETAILS REMARK 245 RECONSTRUCTION METHOD : SINGLE PARTICLE REMARK 245 SPECIMEN TYPE : NULL REMARK 245 REMARK 245 ELECTRON MICROSCOPE SAMPLE REMARK 245 SAMPLE TYPE : PARTICLE REMARK 245 PARTICLE TYPE : POINT REMARK 245 NAME OF SAMPLE : SYNTHETIC HUMAN CLAUDIN-4 REMARK 245 COMPLEX WITH CLOSTRIDIUM REMARK 245 PERFRINGENS ENTEROTOXIN C- REMARK 245 TERMINAL DOMAIN, SFAB COP-2, REMARK 245 AND NANOBODY AGAINST COP-2 REMARK 245 SAMPLE CONCENTRATION (MG ML-1) : 5.00 REMARK 245 SAMPLE SUPPORT DETAILS : ULTRAAUFOIL 1.2/1.3 GRIDS REMARK 245 (QUANTIFOIL) WERE GLOW REMARK 245 DISCHARGED FOR 30 S AT 15 MA IN REMARK 245 A PELCO EASIGLOW (TED PELLA INC) REMARK 245 INSTRUMENT REMARK 245 SAMPLE VITRIFICATION DETAILS : 3.5 MICROL OF COMPLEX WAS REMARK 245 APPLIED ONTO GRIDS AND BLOTTED REMARK 245 FOR 3 S AT 4 DEGREES C UNDER REMARK 245 100 PERCENT HUMIDITY THEN REMARK 245 PLUNGE FROZEN INTO LIQUID REMARK 245 ETHANE COOLED BY LIQUID REMARK 245 NITROGEN. REMARK 245 SAMPLE BUFFER : NULL REMARK 245 PH : 7.40 REMARK 245 SAMPLE DETAILS : ASSEMBLED COMPLEX OF 5 PROTEINS REMARK 245 (FAB IS 2 PROTEINS) EXPRESSED FROM INSECT CELLS AND E COLI REMARK 245 REMARK 245 DATA ACQUISITION REMARK 245 DATE OF EXPERIMENT : NULL REMARK 245 NUMBER OF MICROGRAPHS-IMAGES : 1159 REMARK 245 TEMPERATURE (KELVIN) : NULL REMARK 245 MICROSCOPE MODEL : TFS GLACIOS REMARK 245 DETECTOR TYPE : FEI FALCON IV (4K X 4K) REMARK 245 MINIMUM DEFOCUS (NM) : 400.00 REMARK 245 MAXIMUM DEFOCUS (NM) : 2000.00 REMARK 245 MINIMUM TILT ANGLE (DEGREES) : NULL REMARK 245 MAXIMUM TILT ANGLE (DEGREES) : NULL REMARK 245 NOMINAL CS : NULL REMARK 245 IMAGING MODE : BRIGHT FIELD REMARK 245 ELECTRON DOSE (ELECTRONS NM**-2) : 49.40 REMARK 245 ILLUMINATION MODE : FLOOD BEAM REMARK 245 NOMINAL MAGNIFICATION : 120000 REMARK 245 CALIBRATED MAGNIFICATION : NULL REMARK 245 SOURCE : FIELD EMISSION GUN REMARK 245 ACCELERATION VOLTAGE (KV) : 200 REMARK 245 IMAGING DETAILS : NULL REMARK 247 REMARK 247 ELECTRON MICROSCOPY REMARK 247 THE COORDINATES IN THIS ENTRY WERE GENERATED FROM ELECTRON REMARK 247 MICROSCOPY DATA. PROTEIN DATA BANK CONVENTIONS REQUIRE REMARK 247 THAT CRYST1 AND SCALE RECORDS BE INCLUDED, BUT THE VALUES REMARK 247 ON THESE RECORDS ARE MEANINGLESS EXCEPT FOR THE CALCULATION REMARK 247 OF THE STRUCTURE FACTORS. 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: PENTAMERIC REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, B, H, K, 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 GLY A 186 REMARK 465 GLY A 187 REMARK 465 SER A 188 REMARK 465 LEU A 189 REMARK 465 GLU A 190 REMARK 465 HIS A 191 REMARK 465 HIS A 192 REMARK 465 HIS A 193 REMARK 465 HIS A 194 REMARK 465 HIS A 195 REMARK 465 HIS A 196 REMARK 465 MET B 191 REMARK 465 SER B 192 REMARK 465 THR B 193 REMARK 465 ASP B 194 REMARK 465 ILE B 195 REMARK 465 GLU B 196 REMARK 465 LYS B 197 REMARK 465 GLU B 198 REMARK 465 ILE B 199 REMARK 465 LEU B 200 REMARK 465 ASP B 201 REMARK 465 GLU H 24 REMARK 465 ILE H 25 REMARK 465 SER H 26 REMARK 465 LYS H 253 REMARK 465 SER H 254 REMARK 465 CYS H 255 REMARK 465 ASP H 256 REMARK 465 LYS H 257 REMARK 465 THR H 258 REMARK 465 HIS H 259 REMARK 465 THR H 260 REMARK 465 GLY K 0 REMARK 465 SER K 1 REMARK 465 SER L 25 REMARK 465 CYS L 240 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 ND2 ASN H 130 O TYR H 134 2.02 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 CYS A 54 CA - CB - SG ANGL. DEV. = 13.2 DEGREES REMARK 500 PRO H 186 C - N - CD ANGL. DEV. = -12.8 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 ALA A 153 -60.29 -98.65 REMARK 500 LEU B 271 71.84 52.45 REMARK 500 PHE H 53 -60.80 -109.24 REMARK 500 ASN H 54 160.86 178.02 REMARK 500 ARG H 113 -72.75 -81.63 REMARK 500 ALA H 114 -1.01 -150.73 REMARK 500 ALA H 118 -169.61 -168.70 REMARK 500 TYR H 129 -72.91 -62.81 REMARK 500 ASN H 130 176.36 166.90 REMARK 500 HIS H 133 37.45 71.48 REMARK 500 ALA H 138 -55.84 -127.58 REMARK 500 GLU H 187 109.28 -56.18 REMARK 500 SER H 200 33.54 -142.53 REMARK 500 SER H 211 48.08 -85.19 REMARK 500 SER H 212 13.27 -141.67 REMARK 500 SER K 30 1.00 -64.52 REMARK 500 VAL K 48 -62.01 -126.08 REMARK 500 ARG K 52 157.73 -47.91 REMARK 500 ARG K 53 -163.41 -73.96 REMARK 500 ALA L 68 78.67 54.25 REMARK 500 PRO L 69 152.42 -45.11 REMARK 500 ALA L 76 -5.98 67.61 REMARK 500 SER L 88 11.87 -144.94 REMARK 500 ASP L 107 -159.58 -139.14 REMARK 500 PHE L 108 53.84 70.76 REMARK 500 THR L 110 71.07 54.63 REMARK 500 PRO L 146 -176.78 -69.23 REMARK 500 ALA L 170 148.57 -172.75 REMARK 500 LEU L 227 -165.05 -121.55 REMARK 500 REMARK 500 REMARK: NULL REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: NON-CIS, NON-TRANS REMARK 500 REMARK 500 THE FOLLOWING PEPTIDE BONDS DEVIATE SIGNIFICANTLY FROM BOTH REMARK 500 CIS AND TRANS CONFORMATION. CIS BONDS, IF ANY, ARE LISTED REMARK 500 ON CISPEP RECORDS. TRANS IS DEFINED AS 180 +/- 30 AND REMARK 500 CIS IS DEFINED AS 0 +/- 30 DEGREES. REMARK 500 MODEL OMEGA REMARK 500 TYR H 129 ASN H 130 135.89 REMARK 500 PHE H 185 PRO H 186 -124.35 REMARK 500 SER K 54 GLY K 55 -142.79 REMARK 500 ALA L 109 THR L 110 137.29 REMARK 500 GLN L 225 GLY L 226 146.51 REMARK 500 REMARK 500 REMARK: NULL REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: 7DTM RELATED DB: PDB REMARK 900 RELATED ID: EMD-44479 RELATED DB: EMDB REMARK 900 CRYO-EM STRUCTURE OF SYNTHETIC CLAUDIN-4 COMPLEX WITH CLOSTRIDIUM REMARK 900 PERFRINGENS ENTEROTOXIN C-TERMINAL DOMAIN, SFAB COP-2, AND NANOBODY DBREF 9BEI A 1 196 PDB 9BEI 9BEI 1 196 DBREF 9BEI B 192 319 UNP P01558 ELTB_CLOPF 192 319 DBREF 9BEI H 24 260 PDB 9BEI 9BEI 24 260 DBREF 9BEI K 0 120 PDB 9BEI 9BEI 0 120 DBREF 9BEI L 25 240 PDB 9BEI 9BEI 25 240 SEQADV 9BEI MET B 191 UNP P01558 INITIATING METHIONINE SEQRES 1 A 196 MET SER SER LEU GLU THR PHE ARG GLU ALA ARG ARG LEU SEQRES 2 A 196 ALA ARG GLU GLY LEU GLU LEU VAL ARG GLU ALA ALA ARG SEQRES 3 A 196 LEU PRO MET TRP ARG VAL THR ALA PHE ILE GLY SER ASN SEQRES 4 A 196 ILE VAL THR SER GLN THR ILE TRP GLU GLY LEU TRP MET SEQRES 5 A 196 ASN CYS VAL VAL GLN SER THR GLY GLN MET GLN CYS LYS SEQRES 6 A 196 VAL TYR ASP SER LEU LEU ALA LEU PRO GLU ASP LEU ARG SEQRES 7 A 196 ARG ALA ARG GLU SER PHE GLU ARG ALA ILE GLU VAL ALA SEQRES 8 A 196 GLU LYS ALA LEU GLU LEU LEU GLU ILE GLY ASP PRO ASP SEQRES 9 A 196 SER ASP ALA ILE GLU ASP GLU GLU GLU ARG LEU GLN THR SEQRES 10 A 196 ILE HIS GLU ALA GLY GLU LEU LEU LEU LYS ALA ALA GLU SEQRES 11 A 196 LEU ALA ARG GLU PRO THR GLU ALA ILE ALA ASP ARG ILE SEQRES 12 A 196 ILE GLN ASP PHE TYR ASN PRO LEU VAL ALA SER GLY GLN SEQRES 13 A 196 LYS ARG GLU MET GLY ALA SER LEU ALA LEU ALA ARG ARG SEQRES 14 A 196 GLY ALA GLU LEU LEU GLU GLU ALA GLY ARG LYS LEU LEU SEQRES 15 A 196 GLY LEU GLU GLY GLY SER LEU GLU HIS HIS HIS HIS HIS SEQRES 16 A 196 HIS SEQRES 1 B 129 MET SER THR ASP ILE GLU LYS GLU ILE LEU ASP LEU ALA SEQRES 2 B 129 ALA ALA THR GLU ARG LEU ASN LEU THR ASP ALA LEU ASN SEQRES 3 B 129 SER ASN PRO ALA GLY ASN LEU TYR ASP TRP ARG SER SER SEQRES 4 B 129 ASN SER TYR PRO TRP THR GLN LYS LEU ASN LEU HIS LEU SEQRES 5 B 129 THR ILE THR ALA THR GLY GLN LYS TYR ARG ILE LEU ALA SEQRES 6 B 129 SER LYS ILE VAL ASP PHE ASN ILE TYR SER ASN ASN PHE SEQRES 7 B 129 ASN ASN LEU VAL LYS LEU GLU GLN SER LEU GLY ASP GLY SEQRES 8 B 129 VAL LYS ASP HIS TYR VAL ASP ILE SER LEU ASP ALA GLY SEQRES 9 B 129 GLN TYR VAL LEU VAL MET LYS ALA ASN SER SER TYR SER SEQRES 10 B 129 GLY ASN TYR PRO TYR SER ILE LEU PHE GLN LYS PHE SEQRES 1 H 237 GLU ILE SER GLU VAL GLN LEU VAL GLU SER GLY GLY GLY SEQRES 2 H 237 LEU VAL GLN PRO GLY GLY SER LEU ARG LEU SER CYS ALA SEQRES 3 H 237 ALA SER GLY PHE ASN PHE SER SER SER SER ILE HIS TRP SEQRES 4 H 237 VAL ARG GLN ALA PRO GLY LYS GLY LEU GLU TRP VAL ALA SEQRES 5 H 237 SER ILE SER SER TYR SER GLY TYR THR SER TYR ALA ASP SEQRES 6 H 237 SER VAL LYS GLY ARG PHE THR ILE SER ALA ASP THR SER SEQRES 7 H 237 LYS ASN THR ALA TYR LEU GLN MET ASN SER LEU ARG ALA SEQRES 8 H 237 GLU ASP THR ALA VAL TYR TYR CYS ALA ARG TYR TRP SER SEQRES 9 H 237 TRP TYR ASN SER SER HIS TYR ILE TYR SER ALA LEU ASP SEQRES 10 H 237 TYR TRP GLY GLN GLY THR LEU VAL THR VAL SER SER ALA SEQRES 11 H 237 SER THR LYS GLY PRO SER VAL PHE PRO LEU ALA PRO SER SEQRES 12 H 237 SER LYS SER THR SER GLY GLY THR ALA ALA LEU GLY CYS SEQRES 13 H 237 LEU VAL LYS ASP TYR PHE PRO GLU PRO VAL THR VAL SER SEQRES 14 H 237 TRP ASN SER GLY ALA LEU THR SER GLY VAL HIS THR PHE SEQRES 15 H 237 PRO ALA VAL LEU GLN SER SER GLY LEU TYR SER LEU SER SEQRES 16 H 237 SER VAL VAL THR VAL PRO SER SER SER LEU GLY THR GLN SEQRES 17 H 237 THR TYR ILE CYS ASN VAL ASN HIS LYS PRO SER ASN THR SEQRES 18 H 237 LYS VAL ASP LYS LYS VAL GLU PRO LYS SER CYS ASP LYS SEQRES 19 H 237 THR HIS THR SEQRES 1 K 121 GLY SER VAL GLN LEU GLN GLU SER GLY GLY GLY LEU VAL SEQRES 2 K 121 GLN PRO GLY GLY SER LEU ARG LEU SER CYS ALA ALA SER SEQRES 3 K 121 GLY ARG THR ILE SER ARG TYR ALA MET SER TRP PHE ARG SEQRES 4 K 121 GLN ALA PRO GLY LYS GLU ARG GLU PHE VAL ALA VAL ALA SEQRES 5 K 121 ARG ARG SER GLY ASP GLY ALA PHE TYR ALA ASP SER VAL SEQRES 6 K 121 GLN GLY ARG PHE THR VAL SER ARG ASP ASP ALA LYS ASN SEQRES 7 K 121 THR VAL TYR LEU GLN MET ASN SER LEU LYS PRO GLU ASP SEQRES 8 K 121 THR ALA VAL TYR TYR CYS ALA ILE ASP SER ASP THR PHE SEQRES 9 K 121 TYR SER GLY SER TYR ASP TYR TRP GLY GLN GLY THR GLN SEQRES 10 K 121 VAL THR VAL SER SEQRES 1 L 216 SER ASP ILE GLN MET THR GLN SER PRO SER SER LEU SER SEQRES 2 L 216 ALA SER VAL GLY ASP ARG VAL THR ILE THR CYS ARG ALA SEQRES 3 L 216 SER GLN SER VAL SER SER ALA VAL ALA TRP TYR GLN GLN SEQRES 4 L 216 LYS PRO GLY LYS ALA PRO LYS LEU LEU ILE TYR SER ALA SEQRES 5 L 216 SER SER LEU TYR SER GLY VAL PRO SER ARG PHE SER GLY SEQRES 6 L 216 SER ARG SER GLY THR ASP PHE THR LEU THR ILE SER SER SEQRES 7 L 216 LEU GLN PRO GLU ASP PHE ALA THR TYR TYR CYS GLN GLN SEQRES 8 L 216 SER TYR GLU TRP ALA PRO VAL THR PHE GLY GLN GLY THR SEQRES 9 L 216 LYS VAL GLU ILE LYS ARG THR VAL ALA ALA PRO SER VAL SEQRES 10 L 216 PHE ILE PHE PRO PRO SER ASP SER GLN LEU LYS SER GLY SEQRES 11 L 216 THR ALA SER VAL VAL CYS LEU LEU ASN ASN PHE TYR PRO SEQRES 12 L 216 ARG GLU ALA LYS VAL GLN TRP LYS VAL ASP ASN ALA LEU SEQRES 13 L 216 GLN SER GLY ASN SER GLN GLU SER VAL THR GLU GLN ASP SEQRES 14 L 216 SER LYS ASP SER THR TYR SER LEU SER SER THR LEU THR SEQRES 15 L 216 LEU SER LYS ALA ASP TYR GLU LYS HIS LYS VAL TYR ALA SEQRES 16 L 216 CYS GLU VAL THR HIS GLN GLY LEU SER SER PRO VAL THR SEQRES 17 L 216 LYS SER PHE ASN ARG GLY GLU CYS HELIX 1 AA1 SER A 3 LEU A 27 1 25 HELIX 2 AA2 PRO A 74 GLY A 101 1 28 HELIX 3 AA3 ASP A 110 ASN A 149 1 40 HELIX 4 AA4 GLY A 161 LEU A 184 1 24 HELIX 5 AA5 LEU B 211 ASN B 218 1 8 HELIX 6 AA6 ASN B 267 ASN B 270 5 4 HELIX 7 AA7 SER H 225 GLN H 231 1 7 HELIX 8 AA8 LYS H 240 ASN H 243 5 4 HELIX 9 AA9 LYS K 87 THR K 91 5 5 HELIX 10 AB1 SER L 147 GLY L 154 1 8 HELIX 11 AB2 LYS L 209 HIS L 215 1 7 SHEET 1 AA1 5 GLN A 63 VAL A 66 0 SHEET 2 AA1 5 MET A 52 GLN A 57 -1 N VAL A 55 O GLN A 63 SHEET 3 AA1 5 SER A 43 GLU A 48 -1 N THR A 45 O VAL A 56 SHEET 4 AA1 5 ARG A 31 ILE A 36 -1 N PHE A 35 O GLN A 44 SHEET 5 AA1 5 GLU A 159 MET A 160 -1 O GLU A 159 N VAL A 32 SHEET 1 AA2 5 ARG B 208 ASN B 210 0 SHEET 2 AA2 5 LEU B 238 ILE B 244 1 O HIS B 241 N LEU B 209 SHEET 3 AA2 5 GLY B 294 LYS B 301 -1 O TYR B 296 N LEU B 242 SHEET 4 AA2 5 ASP B 260 SER B 265 -1 N ASP B 260 O LYS B 301 SHEET 5 AA2 5 LYS B 273 LEU B 278 -1 O SER B 277 N PHE B 261 SHEET 1 AA3 4 LEU B 223 ARG B 227 0 SHEET 2 AA3 4 SER B 313 LYS B 318 -1 O ILE B 314 N TRP B 226 SHEET 3 AA3 4 GLN B 249 ALA B 255 -1 N ARG B 252 O GLN B 317 SHEET 4 AA3 4 HIS B 285 LEU B 291 -1 O LEU B 291 N GLN B 249 SHEET 1 AA4 4 GLN H 29 SER H 33 0 SHEET 2 AA4 4 SER H 43 SER H 51 -1 O ALA H 49 N VAL H 31 SHEET 3 AA4 4 THR H 104 ASN H 110 -1 O MET H 109 N LEU H 44 SHEET 4 AA4 4 PHE H 94 ASP H 99 -1 N THR H 95 O GLN H 108 SHEET 1 AA5 5 THR H 84 TYR H 86 0 SHEET 2 AA5 5 LEU H 71 ILE H 77 -1 N SER H 76 O SER H 85 SHEET 3 AA5 5 ILE H 60 GLN H 65 -1 N ARG H 64 O GLU H 72 SHEET 4 AA5 5 VAL H 119 ARG H 124 -1 O VAL H 119 N GLN H 65 SHEET 5 AA5 5 THR H 146 LEU H 147 -1 O THR H 146 N TYR H 120 SHEET 1 AA6 4 SER H 159 LEU H 163 0 SHEET 2 AA6 4 THR H 174 TYR H 184 -1 O GLY H 178 N LEU H 163 SHEET 3 AA6 4 SER H 219 PRO H 224 -1 O VAL H 221 N LEU H 177 SHEET 4 AA6 4 HIS H 203 THR H 204 -1 N HIS H 203 O VAL H 220 SHEET 1 AA7 4 SER H 159 LEU H 163 0 SHEET 2 AA7 4 THR H 174 TYR H 184 -1 O GLY H 178 N LEU H 163 SHEET 3 AA7 4 TYR H 215 LEU H 217 -1 O TYR H 215 N TYR H 184 SHEET 4 AA7 4 VAL H 208 LEU H 209 -1 N VAL H 208 O SER H 216 SHEET 1 AA8 3 THR H 190 ASN H 194 0 SHEET 2 AA8 3 ILE H 234 HIS H 239 -1 O ASN H 238 N THR H 190 SHEET 3 AA8 3 THR H 244 LYS H 249 -1 O VAL H 246 N VAL H 237 SHEET 1 AA9 4 GLN K 3 GLY K 8 0 SHEET 2 AA9 4 LEU K 18 SER K 25 -1 O ALA K 23 N GLN K 5 SHEET 3 AA9 4 THR K 78 MET K 83 -1 O MET K 83 N LEU K 18 SHEET 4 AA9 4 PHE K 68 ASP K 73 -1 N THR K 69 O GLN K 82 SHEET 1 AB1 5 PHE K 59 TYR K 60 0 SHEET 2 AB1 5 ARG K 45 ALA K 51 -1 N VAL K 50 O PHE K 59 SHEET 3 AB1 5 ALA K 33 GLN K 39 -1 N MET K 34 O ALA K 51 SHEET 4 AB1 5 ALA K 92 ASP K 99 -1 O TYR K 95 N PHE K 37 SHEET 5 AB1 5 GLN K 116 VAL K 117 -1 O VAL K 117 N ALA K 92 SHEET 1 AB2 3 MET L 29 GLN L 31 0 SHEET 2 AB2 3 VAL L 44 ALA L 50 -1 O ARG L 49 N THR L 30 SHEET 3 AB2 3 ASP L 95 ILE L 100 -1 O PHE L 96 N CYS L 48 SHEET 1 AB3 2 SER L 35 SER L 37 0 SHEET 2 AB3 2 LYS L 129 GLU L 131 1 O GLU L 131 N LEU L 36 SHEET 1 AB4 5 SER L 78 LEU L 79 0 SHEET 2 AB4 5 LYS L 70 TYR L 74 -1 N TYR L 74 O SER L 78 SHEET 3 AB4 5 TRP L 60 GLN L 62 -1 N TRP L 60 O LEU L 72 SHEET 4 AB4 5 TYR L 111 GLN L 115 -1 O TYR L 112 N TYR L 61 SHEET 5 AB4 5 THR L 123 PHE L 124 -1 O THR L 123 N GLN L 115 SHEET 1 AB5 4 SER L 140 VAL L 141 0 SHEET 2 AB5 4 THR L 155 PHE L 165 -1 O ASN L 163 N SER L 140 SHEET 3 AB5 4 TYR L 199 SER L 208 -1 O SER L 203 N CYS L 160 SHEET 4 AB5 4 SER L 185 VAL L 189 -1 N GLN L 186 O THR L 204 SHEET 1 AB6 3 LYS L 171 TRP L 174 0 SHEET 2 AB6 3 VAL L 217 THR L 223 -1 O THR L 223 N LYS L 171 SHEET 3 AB6 3 VAL L 231 ASN L 236 -1 O PHE L 235 N TYR L 218 SSBOND 1 CYS A 54 CYS A 64 1555 1555 2.05 SSBOND 2 CYS H 48 CYS H 122 1555 1555 2.03 SSBOND 3 CYS H 179 CYS H 235 1555 1555 2.03 SSBOND 4 CYS K 22 CYS K 96 1555 1555 2.03 SSBOND 5 CYS L 48 CYS L 113 1555 1555 2.03 SSBOND 6 CYS L 160 CYS L 220 1555 1555 2.04 CRYST1 1.000 1.000 1.000 90.00 90.00 90.00 P 1 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 1.000000 0.000000 0.000000 0.00000 SCALE2 0.000000 1.000000 0.000000 0.00000 SCALE3 0.000000 0.000000 1.000000 0.00000