HEADER HYDROLASE 27-JUL-05 2AH2 TITLE TRYPANOSOMA CRUZI TRANS-SIALIDASE IN COMPLEX WITH 2,3- TITLE 2 DIFLUOROSIALIC ACID (COVALENT INTERMEDIATE) COMPND MOL_ID: 1; COMPND 2 MOLECULE: TRANS-SIALIDASE; COMPND 3 CHAIN: A; COMPND 4 EC: 3.2.1.18; COMPND 5 ENGINEERED: YES; COMPND 6 MUTATION: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: TRYPANOSOMA CRUZI; SOURCE 3 ORGANISM_COMMON: PROTOZOA; SOURCE 4 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 5 EXPRESSION_SYSTEM_COMMON: BACTERIA; SOURCE 6 EXPRESSION_SYSTEM_STRAIN: TOP10F'; SOURCE 7 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID; SOURCE 8 EXPRESSION_SYSTEM_PLASMID: PTRCHISA KEYWDS TRANSGLYCOSIDASE, COVALENT INTERMEDIATE, TRYPANOSOMA CRUZI, KEYWDS 2 SIALIC ACID EXPDTA X-RAY DIFFRACTION AUTHOR M.F.AMAYA,A.G.WATTS,I.DAMAGER,A.WEHENKEL,T.NGUYEN, AUTHOR 2 A.BUSCHIAZZO,G.PARIS,A.C.FRASCH,S.G.WITHERS,P.M.ALZARI REVDAT 1 23-AUG-05 2AH2 0 SPRSDE 23-AUG-05 2AH2 1S0K JRNL AUTH M.F.AMAYA,A.G.WATTS,I.DAMAGER,A.WEHENKEL,T.NGUYEN, JRNL AUTH 2 A.BUSCHIAZZO,G.PARIS,A.C.FRASCH,S.G.WITHERS, JRNL AUTH 3 P.M.ALZARI JRNL TITL STRUCTURAL INSIGHTS INTO THE CATALYTIC MECHANISM JRNL TITL 2 OF TRYPANOSOMA CRUZI TRANS-SIALIDASE JRNL REF STRUCTURE V. 12 775 2004 JRNL REFN ASTM STRUE6 UK ISSN 0969-2126 REMARK 1 REMARK 2 REMARK 2 RESOLUTION. 1.60 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : REFMAC 5.2.0003 REMARK 3 AUTHORS : MURSHUDOV,VAGIN,DODSON REMARK 3 REMARK 3 REFINEMENT TARGET : ENGH & HUBER REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.60 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 28.40 REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.000 REMARK 3 COMPLETENESS FOR RANGE (%) : 89.3 REMARK 3 NUMBER OF REFLECTIONS : 82504 REMARK 3 REMARK 3 FIT TO DATA USED IN REFINEMENT. REMARK 3 CROSS-VALIDATION METHOD : THROUGHOUT REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM REMARK 3 R VALUE (WORKING + TEST SET) : 0.099 REMARK 3 R VALUE (WORKING SET) : 0.099 REMARK 3 FREE R VALUE : 0.152 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.000 REMARK 3 FREE R VALUE TEST SET COUNT : 4134 REMARK 3 REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN. REMARK 3 TOTAL NUMBER OF BINS USED : 20 REMARK 3 BIN RESOLUTION RANGE HIGH : 1.60 REMARK 3 BIN RESOLUTION RANGE LOW : 1.64 REMARK 3 REFLECTION IN BIN (WORKING SET) : 5424 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 83.63 REMARK 3 BIN R VALUE (WORKING SET) : 0.1030 REMARK 3 BIN FREE R VALUE SET COUNT : 274 REMARK 3 BIN FREE R VALUE : 0.2220 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 ALL ATOMS : 5871 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : NULL REMARK 3 MEAN B VALUE (OVERALL, A**2) : 14.98 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : 0.83000 REMARK 3 B22 (A**2) : -0.50000 REMARK 3 B33 (A**2) : 0.41000 REMARK 3 B12 (A**2) : 0.00000 REMARK 3 B13 (A**2) : 1.16000 REMARK 3 B23 (A**2) : 0.00000 REMARK 3 REMARK 3 ESTIMATED OVERALL COORDINATE ERROR. REMARK 3 ESU BASED ON R VALUE (A): 0.080 REMARK 3 ESU BASED ON FREE R VALUE (A): 0.070 REMARK 3 ESU BASED ON MAXIMUM LIKELIHOOD (A): 0.039 REMARK 3 ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): 2.382 REMARK 3 REMARK 3 CORRELATION COEFFICIENTS. REMARK 3 CORRELATION COEFFICIENT FO-FC : 0.984 REMARK 3 CORRELATION COEFFICIENT FO-FC FREE : 0.968 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES COUNT RMS WEIGHT REMARK 3 BOND LENGTHS REFINED ATOMS (A): 5147 ; 0.019 ; 0.021 REMARK 3 BOND LENGTHS OTHERS (A): 4579 ; 0.001 ; 0.020 REMARK 3 BOND ANGLES REFINED ATOMS (DEGREES): 7004 ; 1.768 ; 1.937 REMARK 3 BOND ANGLES OTHERS (DEGREES): 10667 ; 0.898 ; 3.000 REMARK 3 TORSION ANGLES, PERIOD 1 (DEGREES): 652 ; 7.185 ; 5.000 REMARK 3 TORSION ANGLES, PERIOD 2 (DEGREES): 220 ;33.744 ;23.636 REMARK 3 TORSION ANGLES, PERIOD 3 (DEGREES): 847 ;12.851 ;15.000 REMARK 3 TORSION ANGLES, PERIOD 4 (DEGREES): 32 ;17.281 ;15.000 REMARK 3 CHIRAL-CENTER RESTRAINTS (A**3): 767 ; 0.119 ; 0.200 REMARK 3 GENERAL PLANES REFINED ATOMS (A): 5799 ; 0.009 ; 0.020 REMARK 3 GENERAL PLANES OTHERS (A): 1069 ; 0.001 ; 0.020 REMARK 3 NON-BONDED CONTACTS REFINED ATOMS (A): 922 ; 0.227 ; 0.200 REMARK 3 NON-BONDED CONTACTS OTHERS (A): 4789 ; 0.201 ; 0.200 REMARK 3 NON-BONDED TORSION REFINED ATOMS (A): 2578 ; 0.179 ; 0.200 REMARK 3 NON-BONDED TORSION OTHERS (A): 3149 ; 0.084 ; 0.200 REMARK 3 H-BOND (X...Y) REFINED ATOMS (A): 611 ; 0.207 ; 0.200 REMARK 3 H-BOND (X...Y) OTHERS (A): NULL ; NULL ; NULL REMARK 3 POTENTIAL METAL-ION REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 POTENTIAL METAL-ION OTHERS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY VDW REFINED ATOMS (A): 16 ; 0.228 ; 0.200 REMARK 3 SYMMETRY VDW OTHERS (A): 62 ; 0.271 ; 0.200 REMARK 3 SYMMETRY H-BOND REFINED ATOMS (A): 44 ; 0.381 ; 0.200 REMARK 3 SYMMETRY H-BOND OTHERS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY METAL-ION REFINED ATOMS (A): NULL ; NULL ; NULL REMARK 3 SYMMETRY METAL-ION OTHERS (A): NULL ; NULL ; NULL REMARK 3 REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. COUNT RMS WEIGHT REMARK 3 MAIN-CHAIN BOND REFINED ATOMS (A**2): 4043 ; 2.187 ; 1.500 REMARK 3 MAIN-CHAIN BOND OTHER ATOMS (A**2): 1314 ; 0.655 ; 1.500 REMARK 3 MAIN-CHAIN ANGLE REFINED ATOMS (A**2): 5150 ; 2.657 ; 2.000 REMARK 3 SIDE-CHAIN BOND REFINED ATOMS (A**2): 2336 ; 4.305 ; 3.000 REMARK 3 SIDE-CHAIN ANGLE REFINED ATOMS (A**2): 1854 ; 5.514 ; 4.500 REMARK 3 REMARK 3 ANISOTROPIC THERMAL FACTOR RESTRAINTS. COUNT RMS WEIGHT REMARK 3 RIGID-BOND RESTRAINTS (A**2): 11879 ; 2.413 ;20.000 REMARK 3 SPHERICITY; FREE ATOMS (A**2): 852 ;26.686 ;12.500 REMARK 3 SPHERICITY; BONDED ATOMS (A**2): 9599 ; 6.919 ;12.500 REMARK 3 REMARK 3 NCS RESTRAINTS STATISTICS REMARK 3 NUMBER OF DIFFERENT NCS GROUPS : 0 REMARK 3 REMARK 3 TLS DETAILS REMARK 3 NUMBER OF TLS GROUPS : 0 REMARK 3 REMARK 3 BULK SOLVENT MODELLING. REMARK 3 METHOD USED : BABINET MODEL WITH MASK REMARK 3 PARAMETERS FOR MASK CALCULATION REMARK 3 VDW PROBE RADIUS : 1.20 REMARK 3 ION PROBE RADIUS : 0.80 REMARK 3 SHRINKAGE RADIUS : 0.80 REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: HYDROGENS HAVE BEEN ADDED IN THE REMARK 3 RIDING POSITIONS REMARK 4 REMARK 4 2AH2 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 RCSB033885. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 16-APR-2003 REMARK 200 TEMPERATURE (KELVIN) : 110.0 REMARK 200 PH : 7.50 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : Y REMARK 200 RADIATION SOURCE : ESRF REMARK 200 BEAMLINE : ID14-1 REMARK 200 X-RAY GENERATOR MODEL : NULL REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 0.934 REMARK 200 MONOCHROMATOR : DIAMOND (111), LAUE GEOMETRY, REMARK 200 150 MICRONS THIN REMARK 200 OPTICS : NULL REMARK 200 REMARK 200 DETECTOR TYPE : CCD REMARK 200 DETECTOR MANUFACTURER : ADSC QUANTUM 4 REMARK 200 INTENSITY-INTEGRATION SOFTWARE : DENZO REMARK 200 DATA SCALING SOFTWARE : HKL REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 82813 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.600 REMARK 200 RESOLUTION RANGE LOW (A) : 28.400 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 1.000 REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 89.2 REMARK 200 DATA REDUNDANCY : NULL REMARK 200 R MERGE (I) : 0.04200 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) : 1.60 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.63 REMARK 200 COMPLETENESS FOR SHELL (%) : 83.8 REMARK 200 DATA REDUNDANCY IN SHELL : NULL REMARK 200 R MERGE FOR SHELL (I) : 0.20100 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: FOURIER SYNTHESIS REMARK 200 SOFTWARE USED: AMORE REMARK 200 STARTING MODEL: NULL REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 51.60 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.60 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: PEG 4000, TRIS-HCL, ISOPROPANOL, PH REMARK 280 7.5, VAPOR DIFFUSION, HANGING DROP, TEMPERATURE 291K REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 1 21 1 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -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 64.34500 REMARK 290 SMTRY3 2 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 1 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: A 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 MET A -13 REMARK 465 GLY A -12 REMARK 465 GLY A -11 REMARK 465 SER A -10 REMARK 465 HIS A -9 REMARK 465 HIS A -8 REMARK 465 HIS A -7 REMARK 465 HIS A -6 REMARK 465 HIS A -5 REMARK 465 HIS A -4 REMARK 465 GLY A -3 REMARK 465 MET A -2 REMARK 465 ALA A -1 REMARK 465 SER A 0 REMARK 465 ASP A 400 REMARK 465 PRO A 401 REMARK 465 ALA A 402 REMARK 465 ALA A 403 REMARK 465 SER A 404 REMARK 465 SER A 405 REMARK 465 SER A 406 REMARK 465 GLU A 407 REMARK 465 ARG A 408 REMARK 465 ASP A 634 REMARK 470 REMARK 470 MISSING ATOM REMARK 470 THE FOLLOWING RESIDUES HAVE MISSING ATOMS(M=MODEL NUMBER; REMARK 470 RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE NUMBER; REMARK 470 I=INSERTION CODE): REMARK 470 M RES CSSEQI ATOMS REMARK 470 MET A 633 CB CG SD CE 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 NZ LYS A 215 O HOH 863 1.77 REMARK 500 O HOH 227 O HOH 827 1.80 REMARK 500 O HOH 326 O HOH 859 1.86 REMARK 500 OE2 GLU A 576 O HOH 596 1.89 REMARK 500 O HOH 384 O HOH 825 1.89 REMARK 500 OD2 ASP A 59 O HOH 955 2.02 REMARK 500 O HOH 499 O HOH 920 2.05 REMARK 500 OG SER A 394 O HOH 757 2.10 REMARK 500 O HOH 496 O HOH 761 2.10 REMARK 500 O HOH 666 O HOH 879 2.12 REMARK 500 O HOH 245 O HOH 869 2.13 REMARK 500 O HOH 466 O HOH 680 2.14 REMARK 500 O HOH 576 O HOH 738 2.16 REMARK 500 O HOH 369 O HOH 858 2.17 REMARK 500 O HOH 449 O HOH 682 2.17 REMARK 500 O HOH 355 O HOH 732 2.18 REMARK 500 O HOH 530 O HOH 741 2.19 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 17 O HOH 873 1556 1.40 REMARK 500 O HOH 201 O HOH 954 1554 1.86 REMARK 500 O HOH 362 O HOH 806 2555 2.03 REMARK 500 O HOH 92 O HOH 753 1556 2.10 REMARK 500 O HOH 209 O HOH 431 2555 2.15 REMARK 500 O HOH 718 O HOH 869 2545 2.17 REMARK 500 REMARK 500 GEOMETRY AND STEREOCHEMISTRY REMARK 500 SUBTOPIC: COVALENT BOND LENGTHS 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,2(A3,1X,A1,I4,A1,1X,A4,3X),F6.3) REMARK 500 REMARK 500 EXPECTED VALUES: ENGH AND HUBER, 1991 REMARK 500 REMARK 500 M RES CSSEQI ATM1 RES CSSEQI ATM2 DEVIATION REMARK 500 ILE A 49 CG1 ILE A 49 CD1 -0.143 REMARK 500 LYS A 102 CD LYS A 102 CE -0.186 REMARK 500 LYS A 105 CG LYS A 105 CD -0.125 REMARK 500 MET A 259 CB MET A 259 CG -0.132 REMARK 500 VAL A 420 CB VAL A 420 CG1 -0.132 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: ENGH AND HUBER, 1991 REMARK 500 REMARK 500 M RES CSSEQI ATM1 ATM2 ATM3 REMARK 500 LYS A 12 CD - CE - NZ ANGL. DEV. = 14.9 DEGREES REMARK 500 ARG A 117 CB - CG - CD ANGL. DEV. = 10.8 DEGREES REMARK 500 MET A 259 CB - CG - SD ANGL. DEV. = 16.8 DEGREES 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 753 DISTANCE = 5.09 ANGSTROMS REMARK 525 HOH 818 DISTANCE = 5.39 ANGSTROMS REMARK 600 REMARK 600 HETEROGEN REMARK 600 THE COVALENT SIALO-TYROSINE ADDUCT IS THE PRODUCT OF THE REMARK 600 REACTION OF 2,3-DIFLUOROSIALIC ACID WITH THE ENZYME REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: 2AGS RELATED DB: PDB REMARK 900 CRYSTAL STRUCTURE OF TRYPANOSOMA RANGELI SIALIDASE IN REMARK 900 INCOMPLEX WITH 2-KETO-3-DEOXY-D-GLYCERO-D-GALACTO-2,3_ REMARK 900 DIFLUORO-NONONIC ACID (2,3-DIFLUORO-KDN) REMARK 900 RELATED ID: 2A75 RELATED DB: PDB REMARK 900 CRYSTAL STRUCTURE OF TRYPANOSOMA RANGELI SIALIDASE IN REMARK 900 COMPLEX WITH 2,3- DIFLUOROSIALIC ACID (COVALENT REMARK 900 INTERMEDIATE) REMARK 999 REMARK 999 SEQUENCE REMARK 999 AUTHORS INDICATE THAT THE SEQUENCE IN THE DATABASE IS REMARK 999 INCORRECT DBREF 2AH2 A 1 634 UNP Q26966 Q26966_TRYCR 2 635 SEQADV 2AH2 MET A -13 UNP Q26966 CLONING ARTIFACT SEQADV 2AH2 GLY A -12 UNP Q26966 CLONING ARTIFACT SEQADV 2AH2 GLY A -11 UNP Q26966 CLONING ARTIFACT SEQADV 2AH2 SER A -10 UNP Q26966 CLONING ARTIFACT SEQADV 2AH2 HIS A -9 UNP Q26966 HIS TAG SEQADV 2AH2 HIS A -8 UNP Q26966 HIS TAG SEQADV 2AH2 HIS A -7 UNP Q26966 HIS TAG SEQADV 2AH2 HIS A -6 UNP Q26966 HIS TAG SEQADV 2AH2 HIS A -5 UNP Q26966 HIS TAG SEQADV 2AH2 HIS A -4 UNP Q26966 HIS TAG SEQADV 2AH2 GLY A -3 UNP Q26966 CLONING ARTIFACT SEQADV 2AH2 MET A -2 UNP Q26966 CLONING ARTIFACT SEQADV 2AH2 ALA A -1 UNP Q26966 CLONING ARTIFACT SEQADV 2AH2 SER A 0 UNP Q26966 CLONING ARTIFACT SEQADV 2AH2 PHE A 58 UNP Q26966 ASN 59 ENGINEERED SEQADV 2AH2 THR A 262 UNP Q26966 SER 263 SEE REMARK 999 SEQADV 2AH2 HIS A 476 UNP Q26966 ARG 477 SEE REMARK 999 SEQADV 2AH2 LEU A 484 UNP Q26966 VAL 485 SEE REMARK 999 SEQADV 2AH2 LYS A 495 UNP Q26966 SER 496 ENGINEERED SEQADV 2AH2 GLY A 496 UNP Q26966 VAL 497 ENGINEERED SEQADV 2AH2 LYS A 520 UNP Q26966 GLU 521 ENGINEERED SEQADV 2AH2 VAL A 558 UNP Q26966 GLU 559 SEE REMARK 999 SEQADV 2AH2 GLY A 593 UNP Q26966 ASP 594 ENGINEERED SEQADV 2AH2 ASP A 597 UNP Q26966 ILE 598 ENGINEERED SEQADV 2AH2 ARG A 599 UNP Q26966 HIS 600 ENGINEERED SEQRES 1 A 648 MET GLY GLY SER HIS HIS HIS HIS HIS HIS GLY MET ALA SEQRES 2 A 648 SER LEU ALA PRO GLY SER SER ARG VAL GLU LEU PHE LYS SEQRES 3 A 648 ARG GLN SER SER LYS VAL PRO PHE GLU LYS ASP GLY LYS SEQRES 4 A 648 VAL THR GLU ARG VAL VAL HIS SER PHE ARG LEU PRO ALA SEQRES 5 A 648 LEU VAL ASN VAL ASP GLY VAL MET VAL ALA ILE ALA ASP SEQRES 6 A 648 ALA ARG TYR GLU THR SER PHE ASP ASN SER LEU ILE ASP SEQRES 7 A 648 THR VAL ALA LYS TYR SER VAL ASP ASP GLY GLU THR TRP SEQRES 8 A 648 GLU THR GLN ILE ALA ILE LYS ASN SER ARG ALA SER SER SEQRES 9 A 648 VAL SER ARG VAL VAL ASP PRO THR VAL ILE VAL LYS GLY SEQRES 10 A 648 ASN LYS LEU TYR VAL LEU VAL GLY SER TYR ASN SER SER SEQRES 11 A 648 ARG SER TYR TRP THR SER HIS GLY ASP ALA ARG ASP TRP SEQRES 12 A 648 ASP ILE LEU LEU ALA VAL GLY GLU VAL THR LYS SER THR SEQRES 13 A 648 ALA GLY GLY LYS ILE THR ALA SER ILE LYS TRP GLY SER SEQRES 14 A 648 PRO VAL SER LEU LYS GLU PHE PHE PRO ALA GLU MET GLU SEQRES 15 A 648 GLY MET HIS THR ASN GLN PHE LEU GLY GLY ALA GLY VAL SEQRES 16 A 648 ALA ILE VAL ALA SER ASN GLY ASN LEU VAL TYR PRO VAL SEQRES 17 A 648 GLN VAL THR ASN LYS LYS LYS GLN VAL PHE SER LYS ILE SEQRES 18 A 648 PHE TYR SER GLU ASP GLU GLY LYS THR TRP LYS PHE GLY SEQRES 19 A 648 LYS GLY ARG SER ALA PHE GLY CYS SER GLU PRO VAL ALA SEQRES 20 A 648 LEU GLU TRP GLU GLY LYS LEU ILE ILE ASN THR ARG VAL SEQRES 21 A 648 ASP TYR ARG ARG ARG LEU VAL TYR GLU SER SER ASP MET SEQRES 22 A 648 GLY ASN THR TRP LEU GLU ALA VAL GLY THR LEU SER ARG SEQRES 23 A 648 VAL TRP GLY PRO SER PRO LYS SER ASN GLN PRO GLY SER SEQRES 24 A 648 GLN SER SER PHE THR ALA VAL THR ILE GLU GLY MET ARG SEQRES 25 A 648 VAL MET LEU PHE THR HIS PRO LEU ASN PHE LYS GLY ARG SEQRES 26 A 648 TRP LEU ARG ASP ARG LEU ASN LEU TRP LEU THR ASP ASN SEQRES 27 A 648 GLN ARG ILE TYR ASN VAL GLY GLN VAL SER ILE GLY ASP SEQRES 28 A 648 GLU ASN SER ALA TYR SER SER VAL LEU TYR LYS ASP ASP SEQRES 29 A 648 LYS LEU TYR CYS LEU HIS GLU ILE ASN SER ASN GLU VAL SEQRES 30 A 648 TYR SER LEU VAL PHE ALA ARG LEU VAL GLY GLU LEU ARG SEQRES 31 A 648 ILE ILE LYS SER VAL LEU GLN SER TRP LYS ASN TRP ASP SEQRES 32 A 648 SER HIS LEU SER SER ILE CYS THR PRO ALA ASP PRO ALA SEQRES 33 A 648 ALA SER SER SER GLU ARG GLY CYS GLY PRO ALA VAL THR SEQRES 34 A 648 THR VAL GLY LEU VAL GLY PHE LEU SER HIS SER ALA THR SEQRES 35 A 648 LYS THR GLU TRP GLU ASP ALA TYR ARG CYS VAL ASN ALA SEQRES 36 A 648 SER THR ALA ASN ALA GLU ARG VAL PRO ASN GLY LEU LYS SEQRES 37 A 648 PHE ALA GLY VAL GLY GLY GLY ALA LEU TRP PRO VAL SER SEQRES 38 A 648 GLN GLN GLY GLN ASN GLN ARG TYR HIS PHE ALA ASN HIS SEQRES 39 A 648 ALA PHE THR LEU VAL ALA SER VAL THR ILE HIS GLU VAL SEQRES 40 A 648 PRO LYS GLY ALA SER PRO LEU LEU GLY ALA SER LEU ASP SEQRES 41 A 648 SER SER GLY GLY LYS LYS LEU LEU GLY LEU SER TYR ASP SEQRES 42 A 648 LYS ARG HIS GLN TRP GLN PRO ILE TYR GLY SER THR PRO SEQRES 43 A 648 VAL THR PRO THR GLY SER TRP GLU MET GLY LYS ARG TYR SEQRES 44 A 648 HIS VAL VAL LEU THR MET ALA ASN LYS ILE GLY SER VAL SEQRES 45 A 648 TYR ILE ASP GLY GLU PRO LEU GLU GLY SER GLY GLN THR SEQRES 46 A 648 VAL VAL PRO ASP GLU ARG THR PRO ASP ILE SER HIS PHE SEQRES 47 A 648 TYR VAL GLY GLY TYR LYS ARG SER GLY MET PRO THR ASP SEQRES 48 A 648 SER ARG VAL THR VAL ASN ASN VAL LEU LEU TYR ASN ARG SEQRES 49 A 648 GLN LEU ASN ALA GLU GLU ILE ARG THR LEU PHE LEU SER SEQRES 50 A 648 GLN ASP LEU ILE GLY THR GLU ALA HIS MET ASP HET CL 1001 1 HET FSI A 700 21 HET GOL 1800 6 HET GOL 1801 6 HET GOL 1802 6 HET IPA 1903 4 HETNAM CL CHLORIDE ION HETNAM FSI 5-(ACETYLAMINO)-2,6-ANHYDRO-3,5-DIDEOXY-3- HETNAM 2 FSI FLUORONONONIC ACID HETNAM GOL GLYCEROL HETNAM IPA ISOPROPYL ALCOHOL HETSYN FSI 3-FLUOROSIALIC ACID FORMUL 2 CL CL 1- FORMUL 3 FSI C11 H18 F N O8 FORMUL 4 GOL 3(C3 H8 O3) FORMUL 7 IPA C3 H8 O FORMUL 8 HOH *850(H2 O) HELIX 1 1 TYR A 119 HIS A 123 5 5 HELIX 2 2 THR A 142 LYS A 146 5 5 HELIX 3 3 LYS A 160 PHE A 163 5 4 HELIX 4 4 LEU A 371 ILE A 395 1 25 HELIX 5 5 SER A 467 GLY A 470 5 4 HELIX 6 6 TYR A 475 ASN A 479 5 5 HELIX 7 7 ASN A 613 SER A 623 1 11 HELIX 8 8 THR A 629 MET A 633 5 5 SHEET 1 A 4 SER A 6 PHE A 11 0 SHEET 2 A 4 VAL A 363 ARG A 370 -1 O ARG A 370 N SER A 6 SHEET 3 A 4 LYS A 351 SER A 360 -1 N CYS A 354 O ALA A 369 SHEET 4 A 4 SER A 343 LYS A 348 -1 N LYS A 348 O LYS A 351 SHEET 1 B 2 LYS A 17 LYS A 22 0 SHEET 2 B 2 LYS A 25 VAL A 30 -1 O ARG A 29 N VAL A 18 SHEET 1 C 4 SER A 33 VAL A 42 0 SHEET 2 C 4 VAL A 45 ARG A 53 -1 O VAL A 45 N VAL A 42 SHEET 3 C 4 ILE A 63 SER A 70 -1 O LYS A 68 N ALA A 48 SHEET 4 C 4 GLU A 78 ILE A 83 -1 O GLN A 80 N ALA A 67 SHEET 1 D 3 THR A 148 TRP A 153 0 SHEET 2 D 3 TRP A 129 SER A 141 -1 N THR A 139 O SER A 150 SHEET 3 D 3 VAL A 157 SER A 158 -1 O VAL A 157 N LEU A 133 SHEET 1 E 5 THR A 148 TRP A 153 0 SHEET 2 E 5 TRP A 129 SER A 141 -1 N THR A 139 O SER A 150 SHEET 3 E 5 LYS A 105 TYR A 113 -1 N VAL A 110 O LEU A 132 SHEET 4 E 5 ARG A 93 LYS A 102 -1 N ILE A 100 O TYR A 107 SHEET 5 E 5 GLY A 180 VAL A 181 1 O GLY A 180 N VAL A 99 SHEET 1 F 4 GLU A 166 MET A 167 0 SHEET 2 F 4 MET A 170 GLY A 177 -1 O MET A 170 N MET A 167 SHEET 3 F 4 LEU A 190 ASN A 198 -1 O THR A 197 N ASN A 173 SHEET 4 F 4 ILE A 183 VAL A 184 -1 N ILE A 183 O VAL A 191 SHEET 1 G 5 GLU A 166 MET A 167 0 SHEET 2 G 5 MET A 170 GLY A 177 -1 O MET A 170 N MET A 167 SHEET 3 G 5 LEU A 190 ASN A 198 -1 O THR A 197 N ASN A 173 SHEET 4 G 5 VAL A 203 SER A 210 -1 O PHE A 204 N VAL A 196 SHEET 5 G 5 LYS A 218 PHE A 219 -1 O LYS A 218 N TYR A 209 SHEET 1 H 4 SER A 229 TRP A 236 0 SHEET 2 H 4 LYS A 239 ARG A 245 -1 O ASN A 243 N VAL A 232 SHEET 3 H 4 VAL A 253 SER A 256 -1 O SER A 256 N LEU A 240 SHEET 4 H 4 LEU A 264 GLU A 265 -1 O LEU A 264 N GLU A 255 SHEET 1 I 4 PHE A 289 ILE A 294 0 SHEET 2 I 4 MET A 297 PRO A 305 -1 O VAL A 299 N VAL A 292 SHEET 3 I 4 LEU A 317 THR A 322 -1 O THR A 322 N MET A 300 SHEET 4 I 4 ILE A 327 GLN A 332 -1 O TYR A 328 N LEU A 321 SHEET 1 J 7 ALA A 446 VAL A 449 0 SHEET 2 J 7 GLY A 452 PHE A 455 -1 O LYS A 454 N GLU A 447 SHEET 3 J 7 VAL A 600 TYR A 608 -1 O VAL A 600 N PHE A 455 SHEET 4 J 7 ALA A 481 ILE A 490 -1 N THR A 483 O TYR A 608 SHEET 5 J 7 ARG A 544 ALA A 552 -1 O VAL A 547 N ALA A 486 SHEET 6 J 7 ILE A 555 ILE A 560 -1 O TYR A 559 N VAL A 548 SHEET 7 J 7 GLU A 563 PRO A 564 -1 O GLU A 563 N ILE A 560 SHEET 1 K13 TRP A 524 TYR A 528 0 SHEET 2 K13 LYS A 512 ASP A 519 -1 N SER A 517 O GLN A 525 SHEET 3 K13 ALA A 497 SER A 504 -1 N LEU A 501 O LEU A 516 SHEET 4 K13 HIS A 583 VAL A 586 -1 O HIS A 583 N SER A 504 SHEET 5 K13 GLY A 461 PRO A 465 -1 N TRP A 464 O PHE A 584 SHEET 6 K13 ALA A 441 ALA A 444 -1 N SER A 442 O LEU A 463 SHEET 7 K13 GLU A 431 ASP A 434 -1 N TRP A 432 O ALA A 441 SHEET 8 K13 LEU A 419 ALA A 427 -1 N SER A 426 O GLU A 433 SHEET 9 K13 VAL A 600 TYR A 608 -1 O LEU A 607 N VAL A 420 SHEET 10 K13 ALA A 481 ILE A 490 -1 N THR A 483 O TYR A 608 SHEET 11 K13 ARG A 544 ALA A 552 -1 O VAL A 547 N ALA A 486 SHEET 12 K13 ILE A 555 ILE A 560 -1 O TYR A 559 N VAL A 548 SHEET 13 K13 GLN A 570 THR A 571 -1 O GLN A 570 N GLY A 556 SSBOND 1 CYS A 396 CYS A 410 LINK OH TYR A 342 C2 FSI A 700 CRYST1 54.149 128.690 54.298 90.00 108.74 90.00 P 1 21 1 2 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 0.018468 0.000000 0.006265 0.00000 SCALE2 0.000000 0.007771 0.000000 0.00000 SCALE3 0.000000 0.000000 0.019448 0.00000