HEADER HYDROLASE INHIBITOR 08-APR-02 1LD6
TITLE STRUCTURE OF BPTI_8A MUTANT
COMPND MOL_ID: 1;
COMPND 2 MOLECULE: PANCREATIC TRYPSIN INHIBITOR;
COMPND 3 CHAIN: A;
COMPND 4 ENGINEERED: YES;
COMPND 5 MUTATION: YES
SOURCE MOL_ID: 1;
SOURCE 2 ORGANISM_SCIENTIFIC: BOS TAURUS;
SOURCE 3 ORGANISM_COMMON: CATTLE;
SOURCE 4 ORGANISM_TAXID: 9913;
SOURCE 5 EXPRESSION_SYSTEM: ESCHERICHIA COLI;
SOURCE 6 EXPRESSION_SYSTEM_TAXID: 562;
SOURCE 7 EXPRESSION_SYSTEM_STRAIN: BL21 (DE3) PLYSS;
SOURCE 8 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID;
SOURCE 9 EXPRESSION_SYSTEM_PLASMID: PAED4
KEYWDS BPTI, KUNITZ FOLD, HYDROLASE INHIBITOR
EXPDTA SOLUTION NMR
NUMMDL 10
AUTHOR T.CIERPICKI,J.OTLEWSKI
REVDAT 3 27-OCT-21 1LD6 1 REMARK SEQADV
REVDAT 2 24-FEB-09 1LD6 1 VERSN
REVDAT 1 11-SEP-02 1LD6 0
JRNL AUTH T.CIERPICKI,J.OTLEWSKI
JRNL TITL NMR STRUCTURES OF TWO VARIANTS OF BOVINE PANCREATIC TRYPSIN
JRNL TITL 2 INHIBITOR (BPTI) REVEAL UNEXPECTED INFLUENCE OF MUTATIONS ON
JRNL TITL 3 PROTEIN STRUCTURE AND STABILITY.
JRNL REF J.MOL.BIOL. V. 321 647 2002
JRNL REFN ISSN 0022-2836
JRNL PMID 12206780
JRNL DOI 10.1016/S0022-2836(02)00620-4
REMARK 1
REMARK 1 REFERENCE 1
REMARK 1 AUTH O.BUCZEK,K.KOSCIELSKA-KASPRZAK,D.KROWARSCH,M.DADLEZ,
REMARK 1 AUTH 2 M.OTLEWSKI
REMARK 1 TITL ANALYSIS OF SERINE PROTEINASE-INHIBITOR INTERACTION BY
REMARK 1 TITL 2 ALANINE SHAVING
REMARK 1 REF PROTEIN SCI. V. 11 806 2002
REMARK 1 REFN ISSN 0961-8368
REMARK 1 DOI 10.1110/PS.3510102
REMARK 2
REMARK 2 RESOLUTION. NOT APPLICABLE.
REMARK 3
REMARK 3 REFINEMENT.
REMARK 3 PROGRAM : NMRPIPE 1.8, CNS 1.0
REMARK 3 AUTHORS : DELAGLIO, F.; GRZESIEK, S.; VUISTER, G.; ZHU, G.;
REMARK 3 PFEIFER, J.; BAX, A. (NMRPIPE), BRUNGER, A.T.;
REMARK 3 ADAMS, P.D.; CLORE, G.M.; DELANO, W.L., GROS,P.;
REMARK 3 GROSSE-KUNSTLEVE, R.W.; JIANG, J.S.; KUSZEWSKI, J.;
REMARK 3 NILGES, M.; PANNU, N.S.; READ, R.J.; RICE, L.M.;
REMARK 3 SIMONSON, T.; WARREN, G.L. (CNS)
REMARK 3
REMARK 3 OTHER REFINEMENT REMARKS: NULL
REMARK 4
REMARK 4 1LD6 COMPLIES WITH FORMAT V. 3.15, 01-DEC-08
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 10-APR-02.
REMARK 100 THE DEPOSITION ID IS D_1000015852.
REMARK 210
REMARK 210 EXPERIMENTAL DETAILS
REMARK 210 EXPERIMENT TYPE : NMR
REMARK 210 TEMPERATURE (KELVIN) : 298
REMARK 210 PH : 2.9
REMARK 210 IONIC STRENGTH : 0
REMARK 210 PRESSURE : AMBIENT
REMARK 210 SAMPLE CONTENTS : 3MM BPTI_8A
REMARK 210
REMARK 210 NMR EXPERIMENTS CONDUCTED : 2D TOCSY; 2D NOESY; DQF-COSY
REMARK 210 SPECTROMETER FIELD STRENGTH : 500 MHZ
REMARK 210 SPECTROMETER MODEL : UNITYPLUS; DRX
REMARK 210 SPECTROMETER MANUFACTURER : VARIAN; BRUKER
REMARK 210
REMARK 210 STRUCTURE DETERMINATION.
REMARK 210 SOFTWARE USED : SPARKY 3.95, DYANA 1.5
REMARK 210 METHOD USED : SIMULATED ANNEALING, TORSION
REMARK 210 ANGLE DYNAMICS
REMARK 210
REMARK 210 CONFORMERS, NUMBER CALCULATED : 58
REMARK 210 CONFORMERS, NUMBER SUBMITTED : 10
REMARK 210 CONFORMERS, SELECTION CRITERIA : STRUCTURES WITH THE LOWEST
REMARK 210 ENERGY,TARGET FUNCTION
REMARK 210
REMARK 210 BEST REPRESENTATIVE CONFORMER IN THIS ENSEMBLE : 1
REMARK 210
REMARK 210 REMARK: STRUCTURE WAS DETERMINED BASED ON HOMONUCLEAR TOCSY/NOESY
REMARK 210 TECHNIQUES USING AUTOMATIC ASSIGNMENT IN NOAH/DYANA PROGRAM
REMARK 215
REMARK 215 NMR STUDY
REMARK 215 THE COORDINATES IN THIS ENTRY WERE GENERATED FROM SOLUTION
REMARK 215 NMR DATA. PROTEIN DATA BANK CONVENTIONS REQUIRE THAT
REMARK 215 CRYST1 AND SCALE RECORDS BE INCLUDED, BUT THE VALUES ON
REMARK 215 THESE RECORDS ARE MEANINGLESS.
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: MONOMERIC
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 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 1 ALA A 13 97.96 -161.89
REMARK 500 1 ALA A 17 44.34 -161.54
REMARK 500 1 ALA A 19 45.26 -79.48
REMARK 500 1 ALA A 27 -1.94 -141.43
REMARK 500 1 THR A 32 31.80 -83.23
REMARK 500 1 PHE A 33 159.63 65.48
REMARK 500 1 TYR A 35 -165.60 43.75
REMARK 500 2 ALA A 11 75.13 -103.97
REMARK 500 2 ALA A 17 41.06 -160.49
REMARK 500 2 ALA A 18 80.36 -175.79
REMARK 500 2 ALA A 19 -55.14 -163.40
REMARK 500 2 PHE A 33 106.16 -46.39
REMARK 500 2 LYS A 41 -79.03 -62.90
REMARK 500 3 ARG A 15 88.41 -154.48
REMARK 500 3 ALA A 17 47.05 -159.16
REMARK 500 3 TYR A 35 65.05 -64.57
REMARK 500 3 LYS A 41 -77.15 -60.97
REMARK 500 4 PRO A 9 170.22 -41.46
REMARK 500 4 ALA A 13 36.00 -86.67
REMARK 500 4 ALA A 16 95.47 64.99
REMARK 500 4 ALA A 17 50.94 -162.57
REMARK 500 4 ALA A 19 -48.80 -165.00
REMARK 500 4 ALA A 34 -159.28 -166.07
REMARK 500 4 TYR A 35 140.86 -36.91
REMARK 500 4 ALA A 37 91.61 -162.63
REMARK 500 4 ARG A 42 -82.76 -84.57
REMARK 500 5 ALA A 13 35.06 70.63
REMARK 500 5 ALA A 16 101.36 71.69
REMARK 500 5 ALA A 17 44.92 -163.02
REMARK 500 5 ALA A 19 21.70 -162.64
REMARK 500 5 ALA A 37 -178.97 -69.74
REMARK 500 5 LYS A 41 -80.78 -59.77
REMARK 500 6 ARG A 15 81.53 -160.44
REMARK 500 6 ALA A 17 62.83 71.04
REMARK 500 6 ALA A 19 73.41 32.42
REMARK 500 6 LYS A 41 -68.28 91.19
REMARK 500 7 ALA A 13 94.75 56.49
REMARK 500 7 ALA A 16 90.36 80.03
REMARK 500 7 ALA A 17 64.90 -160.85
REMARK 500 7 ALA A 19 92.13 56.59
REMARK 500 7 TYR A 35 64.13 -66.64
REMARK 500 7 LYS A 41 -75.35 -63.78
REMARK 500 7 ASN A 44 115.65 74.00
REMARK 500 8 PRO A 9 -162.74 -74.94
REMARK 500 8 ALA A 17 60.33 -160.60
REMARK 500 8 ALA A 19 -39.67 -164.21
REMARK 500 8 LYS A 41 -81.24 -62.50
REMARK 500 8 THR A 54 -70.65 -80.07
REMARK 500 9 ALA A 16 119.14 67.19
REMARK 500 9 ALA A 17 44.70 -161.78
REMARK 500
REMARK 500 THIS ENTRY HAS 59 RAMACHANDRAN OUTLIERS.
REMARK 500
REMARK 500 REMARK: NULL
DBREF 1LD6 A 1 58 UNP P00974 BPT1_BOVIN 36 93
SEQADV 1LD6 ALA A 11 UNP P00974 THR 46 ENGINEERED MUTATION
SEQADV 1LD6 ALA A 13 UNP P00974 PRO 48 ENGINEERED MUTATION
SEQADV 1LD6 ARG A 15 UNP P00974 LYS 50 CONFLICT
SEQADV 1LD6 ALA A 17 UNP P00974 ARG 52 ENGINEERED MUTATION
SEQADV 1LD6 ALA A 18 UNP P00974 ILE 53 ENGINEERED MUTATION
SEQADV 1LD6 ALA A 19 UNP P00974 ILE 54 ENGINEERED MUTATION
SEQADV 1LD6 ALA A 34 UNP P00974 VAL 69 ENGINEERED MUTATION
SEQADV 1LD6 ALA A 37 UNP P00974 GLY 72 ENGINEERED MUTATION
SEQADV 1LD6 ALA A 39 UNP P00974 ARG 74 ENGINEERED MUTATION
SEQADV 1LD6 LEU A 52 UNP P00974 MET 87 CONFLICT
SEQRES 1 A 58 ARG PRO ASP PHE CYS LEU GLU PRO PRO TYR ALA GLY ALA
SEQRES 2 A 58 CYS ARG ALA ALA ALA ALA ARG TYR PHE TYR ASN ALA LYS
SEQRES 3 A 58 ALA GLY LEU CYS GLN THR PHE ALA TYR GLY ALA CYS ALA
SEQRES 4 A 58 ALA LYS ARG ASN ASN PHE LYS SER ALA GLU ASP CYS LEU
SEQRES 5 A 58 ARG THR CYS GLY GLY ALA
HELIX 1 1 PRO A 2 GLU A 7 5 6
HELIX 2 2 SER A 47 CYS A 55 1 9
SHEET 1 A 2 PHE A 22 TYR A 23 0
SHEET 2 A 2 CYS A 30 GLN A 31 -1 O GLN A 31 N PHE A 22
SSBOND 1 CYS A 5 CYS A 55 1555 1555 2.03
SSBOND 2 CYS A 14 CYS A 38 1555 1555 2.03
SSBOND 3 CYS A 30 CYS A 51 1555 1555 2.03
CRYST1 1.000 1.000 1.000 90.00 90.00 90.00 P 1 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
MODEL 1
(ATOM LINES ARE NOT SHOWN.)
END
