HEADER COMPLEX(SERINE PROTEINASE-INHIBITOR) 03-JUN-88 1CSE 1CSE 3 COMPND SUBTILISIN CARLSBERG (E.C.3.4.21.62) (COMMERCIAL PRODUCT 1CSEC 1 COMPND 1 FROM SERRA, HEIDELBERG CALLED SUBTILISIN NAGARSE) COMPLEX 1CSE 5 COMPND 2 WITH EGLIN-C 1CSE 6 SOURCE (BACILLUS $SUBTILIS) AND LEECH (HIRUDO $MEDICINALIS) 1CSE 7 AUTHOR W.BODE 1CSE 8 REVDAT 4 15-JAN-95 1CSEC 1 COMPND HET 1CSEC 2 REVDAT 3 15-OCT-89 1CSEB 1 SEQRES 1CSEB 1 REVDAT 2 19-APR-89 1CSEA 1 REMARK 1CSEA 1 REVDAT 1 16-JUL-88 1CSE 0 1CSE 9 REMARK 1 1CSE 10 REMARK 1 REFERENCE 1 1CSE 11 REMARK 1 AUTH W.BODE,E.PAPAMOKOS,D.MUSIL 1CSE 12 REMARK 1 TITL THE HIGH-RESOLUTION X-RAY CRYSTAL STRUCTURE OF THE 1CSE 13 REMARK 1 TITL 2 COMPLEX FORMED BETWEEN SUBTILISIN CARLSBERG AND 1CSE 14 REMARK 1 TITL 3 EGLIN $C, AN ELASTASE INHIBITOR FROM THE LEECH 1CSE 15 REMARK 1 TITL 4 HIRUDO $MEDICINALIS. STRUCTURAL ANALYSIS, 1CSE 16 REMARK 1 TITL 5 SUBTILISIN STRUCTURE AND INTERFACE GEOMETRY 1CSE 17 REMARK 1 REF EUR.J.BIOCHEM. V. 166 673 1987 1CSE 18 REMARK 1 REFN ASTM EJBCAI IX ISSN 0014-2956 262 1CSE 19 REMARK 1 REFERENCE 2 1CSE 20 REMARK 1 AUTH W.BODE,E.PAPAMOKOS,D.MUSIL,U.SEEMUELLER,H.FRITZ 1CSE 21 REMARK 1 TITL REFINED 1.2 ANGSTROMS CRYSTAL STRUCTURE OF THE 1CSE 22 REMARK 1 TITL 2 COMPLEX FORMED BETWEEN SUBTILISIN CARLSBERG AND THE 1CSE 23 REMARK 1 TITL 3 INHIBITOR EGLIN $C. MOLECULAR STRUCTURE OF EGLIN 1CSE 24 REMARK 1 TITL 4 AND ITS DETAILED INTERACTION WITH SUBTILISIN 1CSE 25 REMARK 1 REF /EMBO$ J. V. 5 813 1986 1CSE 26 REMARK 1 REFN ASTM EMJODG UK ISSN 0261-4189 897 1CSE 27 REMARK 1 REFERENCE 3 1CSE 28 REMARK 1 AUTH C.A.MC*PHALEN,H.P.SCHNEBLI,M.N.G.JAMES 1CSE 29 REMARK 1 TITL CRYSTAL AND MOLECULAR STRUCTURE OF THE INHIBITOR 1CSE 30 REMARK 1 TITL 2 EGLIN FROM LEECHES IN COMPLEX WITH SUBTILISIN 1CSE 31 REMARK 1 TITL 3 CARLSBERG 1CSE 32 REMARK 1 REF /FEBS$ LETT. V. 188 55 1985 1CSE 33 REMARK 1 REFN ASTM FEBLAL NE ISSN 0014-5793 165 1CSE 34 REMARK 2 1CSE 35 REMARK 2 RESOLUTION. 1.2 ANGSTROMS. 1CSE 36 REMARK 3 1CSE 37 REMARK 3 REFINEMENT. BY THE RECIPROCAL SPACE REFINEMENT PROCEDURE 1CSE 38 REMARK 3 WITH ENERGY RESTRAINTS OF A. JACK AND M. LEVITT (PROGRAM 1CSE 39 REMARK 3 *EREF*). THE R VALUE FOR 44500 REFLECTIONS IN THE 1CSE 40 REMARK 3 RESOLUTION RANGE 10.0 TO 1.2 ANGSTROMS (OMITTING 1.5 PER 1CSE 41 REMARK 3 CENT OF MAINLY WEAK REFLECTIONS FOR WHICH 1CSE 42 REMARK 3 2.0 * ABS(ABS(FOBS)-ABS(FCALC)) / (ABS(FOBS)+ABS(FCALC)) 1CSEA 2 REMARK 3 .GT. 1.2 IS 0.178 FOR K=6.0*10E-4 (A FACTOR RELATING THE 1CSEA 3 REMARK 3 CRYSTALLOGRAPHIC AND ENERGY TERMS IN EREF). THE RMS 1CSE 45 REMARK 3 DEVIATION FROM IDEALITY OF THE BOND DISTANCES IS 0.022 1CSE 46 REMARK 3 ANGSTROMS. THE RMS DEVIATION FROM IDEALITY OF THE BOND 1CSE 47 REMARK 3 ANGLES IS 2.48 DEGREES. ALSO SEE REMARK 5. ATOMS WITH 1CSE 48 REMARK 3 THERMAL FACTORS WHICH CALCULATE LESS THAN 3.00 ARE 1CSE 49 REMARK 3 ASSIGNED THIS VALUE. THIS IS THE LOWEST VALUE ALLOWED BY 1CSE 50 REMARK 3 THE REFINEMENT PROGRAM. 1CSE 51 REMARK 4 1CSE 52 REMARK 4 THE SEQUENCE NUMBERING USED FOR SUBTILISIN IS BASED ON THAT 1CSE 53 REMARK 4 OF SUBTILISIN BPN(PRIME) WITH A SINGLE DELETION AT RESIDUE 1CSE 54 REMARK 4 56. THE EGLIN NOMENCLATURE (70 RESIDUES) IS USED. THE 1CSE 55 REMARK 4 EGLIN CHAIN CAN ONLY BE TRACED FROM LYS 8 ONWARDS. 1CSE 56 REMARK 4 ACCORDING TO PAGE SOME N-TERMINAL RESIDUES (MOST PROBABLY 1CSE 57 REMARK 4 THE FIRST SIX RESIDUES) ARE CLEAVED OFF IN THE COURSE OF 1CSE 58 REMARK 4 ACTIVATION. SUBTILISIN HAS BEEN ASSIGNED CHAIN IDENTIFIER 1CSE 59 REMARK 4 *E* AND EGLIN C HAS BEEN ASSIGNED CHAIN IDENTIFIER *I*. 1CSE 60 REMARK 5 1CSE 61 REMARK 5 RESIDUE THR 211 IS A CIS THREONINE. IT WAS ORIGINALLY 1CSE 62 REMARK 5 MODELLED AS A TRANS CONFORMER. AS OUTLINED IN REFERENCE 1 1CSE 63 REMARK 5 ABOVE, HOWEVER, THE MAIN CHAIN ANGLES OF THIS RESIDUE WERE 1CSE 64 REMARK 5 OUTSIDE THE ALLOWED REGIONS AND THE FIT TO THE ELECTRON 1CSE 65 REMARK 5 DENSITY WAS STILL INSUFFICIENT. THE CIS CONFORMER GIVEN IN 1CSE 66 REMARK 5 THIS ENTRY FITS MUCH BETTER. THE MODIFIED MODEL HAS BEEN 1CSE 67 REMARK 5 SUBJECTED TO TWO FURTHER MINICYCLES OF POSITIONAL AND B 1CSE 68 REMARK 5 FACTOR REFINEMENT WITHOUT GROSS CONFORMATION CHANGES AND 1CSE 69 REMARK 5 WITHOUT AFFECTING THE R VALUE. TWO SITES PROBABLY OCCUPIED 1CSE 70 REMARK 5 BY CALCIUM IONS AND 432 SOLVENT MOLECULES WERE LOCATED. 1CSE 71 REMARK 5 FOR THESE 434 NON-PROTEIN ATOMS REFINED INDIVIDUAL 1CSE 72 REMARK 5 OCCUPANCIES ARE GIVEN. 1CSE 73 REMARK 6 1CSE 74 REMARK 6 SITE *ACT* IS THE ENZYME CATALYTIC SITE. THE INHIBITOR 1CSE 75 REMARK 6 REACTIVE SITE BOND IS LOCATED BETWEEN RESIDUES LEU I 59 1CSE 76 REMARK 6 AND ASP I 60 (SITE *RSB*). 1CSE 77 REMARK 7 1CSEA 4 REMARK 7 CORRECTION. CORRECT FORMULA GIVEN IN REMARK 3. 19-APR-89. 1CSEA 5 REMARK 8 1CSEB 2 REMARK 8 CORRECTION. CORRECT NUMBER OF RESIDUES ON SEQRES RECORDS. 1CSEB 3 REMARK 8 CORRECT ORDER OF RESIDUES ON SEQRES RECORD. 15-OCT-89. 1CSEB 4 REMARK 9 1CSEC 3 REMARK 9 CORRECTION. DELETE CHAIN IDENTIFIER FROM HET RECORDS. 1CSEC 4 REMARK 9 CORRECT E.C. CODE ON COMPND RECORD. 15-JAN-95. 1CSEC 5 SEQRES 1 E 274 ALA GLN THR VAL PRO TYR GLY ILE PRO LEU ILE LYS ALA 1CSEB 5 SEQRES 2 E 274 ASP LYS VAL GLN ALA GLN GLY PHE LYS GLY ALA ASN VAL 1CSEB 6 SEQRES 3 E 274 LYS VAL ALA VAL LEU ASP THR GLY ILE GLN ALA SER HIS 1CSEB 7 SEQRES 4 E 274 PRO ASP LEU ASN VAL VAL GLY GLY ALA SER PHE VAL ALA 1CSEB 8 SEQRES 5 E 274 GLY GLU ALA TYR ASN THR ASP GLY ASN GLY HIS GLY THR 1CSEB 9 SEQRES 6 E 274 HIS VAL ALA GLY THR VAL ALA ALA LEU ASP ASN THR THR 1CSEB 10 SEQRES 7 E 274 GLY VAL LEU GLY VAL ALA PRO SER VAL SER LEU TYR ALA 1CSEB 11 SEQRES 8 E 274 VAL LYS VAL LEU ASN SER SER GLY SER GLY SER TYR SER 1CSEB 12 SEQRES 9 E 274 GLY ILE VAL SER GLY ILE GLU TRP ALA THR THR ASN GLY 1CSEB 13 SEQRES 10 E 274 MET ASP VAL ILE ASN MET SER LEU GLY GLY ALA SER GLY 1CSEB 14 SEQRES 11 E 274 SER THR ALA MET LYS GLN ALA VAL ASP ASN ALA TYR ALA 1CSEB 15 SEQRES 12 E 274 ARG GLY VAL VAL VAL VAL ALA ALA ALA GLY ASN SER GLY 1CSEB 16 SEQRES 13 E 274 ASN SER GLY SER THR ASN THR ILE GLY TYR PRO ALA LYS 1CSEB 17 SEQRES 14 E 274 TYR ASP SER VAL ILE ALA VAL GLY ALA VAL ASP SER ASN 1CSEB 18 SEQRES 15 E 274 SER ASN ARG ALA SER PHE SER SER VAL GLY ALA GLU LEU 1CSEB 19 SEQRES 16 E 274 GLU VAL MET ALA PRO GLY ALA GLY VAL TYR SER THR TYR 1CSEB 20 SEQRES 17 E 274 PRO THR ASN THR TYR ALA THR LEU ASN GLY THR SER MET 1CSEB 21 SEQRES 18 E 274 ALA SER PRO HIS VAL ALA GLY ALA ALA ALA LEU ILE LEU 1CSEB 22 SEQRES 19 E 274 SER LYS HIS PRO ASN LEU SER ALA SER GLN VAL ARG ASN 1CSEB 23 SEQRES 20 E 274 ARG LEU SER SER THR ALA THR TYR LEU GLY SER SER PHE 1CSEB 24 SEQRES 21 E 274 TYR TYR GLY LYS GLY LEU ILE ASN VAL GLU ALA ALA ALA 1CSEB 25 SEQRES 22 E 274 GLN 1CSEB 26 SEQRES 1 I 71 ACE THR GLU PHE GLY SER GLU LEU LYS SER PHE PRO GLU 1CSE 100 SEQRES 2 I 71 VAL VAL GLY LYS THR VAL ASP GLN ALA ARG GLU TYR PHE 1CSE 101 SEQRES 3 I 71 THR LEU HIS TYR PRO GLN TYR ASN VAL TYR PHE LEU PRO 1CSE 102 SEQRES 4 I 71 GLU GLY SER PRO VAL THR LEU ASP LEU ARG TYR ASN ARG 1CSEB 27 SEQRES 5 I 71 VAL ARG VAL PHE TYR ASN PRO GLY THR ASN VAL VAL ASN 1CSE 104 SEQRES 6 I 71 HIS VAL PRO HIS VAL GLY 1CSE 105 FTNOTE 1 1CSE 106 FTNOTE 1 RESIDUE PRO 168 IS A CIS PROLINE. 1CSE 107 FTNOTE 2 1CSE 108 FTNOTE 2 RESIDUE THR 211 IS A CIS THREONINE. 1CSE 109 FTNOTE 3 1CSE 110 FTNOTE 3 SEE REMARK 3. 1CSE 111 HET CA 401 1 CALCIUM ++ ION 1CSEC 6 HET CA 430 1 CALCIUM ++ ION 1CSEC 7 FORMUL 3 CA 2(CA1 ++) 1CSE 114 FORMUL 4 HOH *432(H2 O1) 1CSE 115 HELIX 1 AE TYR E 6 ILE E 11 1 1CSE 116 HELIX 2 BE LYS E 12 GLN E 19 1 1CSE 117 HELIX 3 CE GLY E 63 ALA E 74 1 1CSE 118 HELIX 4 DE SER E 103 ASN E 117 1 1CSE 119 HELIX 5 EE SER E 132 GLY E 146 1 1CSE 120 HELIX 6 EF THR E 220 HIS E 238 1 WITH PERTURBATION AT SER 221 1CSE 121 HELIX 7 GE SER E 242 THR E 253 1 1CSE 122 HELIX 9 HE ASN E 269 ALA E 274 1 1CSE 123 HELIX 10 IA PHE I 10 VAL I 14 5 1CSE 124 HELIX 11 IB THR I 17 TYR I 29 1 1CSE 125 SHEET 1 S1E 8 ASN E 43 PHE E 50 0 1CSE 126 SHEET 2 S1E 8 SER E 89 VAL E 95 1 1CSE 127 SHEET 3 S1E 8 VAL E 26 ASP E 32 1 1CSE 128 SHEET 4 S1E 8 ASP E 120 MET E 124 1 1CSE 129 SHEET 5 S1E 8 VAL E 148 ALA E 153 1 1CSE 130 SHEET 6 S1E 8 ILE E 175 VAL E 180 1 1CSE 131 SHEET 7 S1E 8 GLU E 197 GLY E 202 1 1CSE 132 SHEET 8 S1E 8 LYS E 265 ILE E 268 1 1CSE 133 SHEET 1 S1I 4 LYS I 8 PHE I 10 0 1CSE 134 SHEET 2 S1I 4 HIS I 65 GLY I 70 -1 1CSE 135 SHEET 3 S1I 4 ARG I 51 TYR I 56 -1 1CSE 136 SHEET 4 S1I 4 ASN I 33 LEU I 37 1 1CSE 137 TURN 1 1E PRO E 5 ILE E 8 TYPE III 1CSE 138 TURN 2 2E PRO E 9 LYS E 12 TYPE I 1CSE 139 TURN 3 3E GLN E 17 GLY E 20 TYPE III 1CSE 140 TURN 4 4E GLY E 23 VAL E 26 TYPE II 1CSE 141 TURN 5 5E GLN E 36 HIS E 39 TYPE I 1CSE 142 TURN 6 6E HIS E 39 LEU E 42 TYPE I 1CSE 143 TURN 7 7E VAL E 51 GLU E 54 TYPE II 1CSE 144 TURN 8 8E ALA E 55 THR E 59 TYPE I 1CSE 145 TURN 9 9E ALA E 85 VAL E 88 TYPE I 1CSE 146 TURN 10 10E ASN E 97 GLY E 100 TYPE I 1CSE 147 TURN 11 11E TYR E 143 GLY E 146 TYPE III 1CSE 148 TURN 12 12E SER E 159 THR E 162 TYPE II(PRIME) 1CSE 149 TURN 13 13E PRO E 168 TYR E 171 TYPE III 1CSE 150 TURN 14 14E TYR E 171 VAL E 174 TYPE I 1CSE 151 TURN 15 15E ASP E 181 SER E 184 TYPE I 1CSE 152 TURN 16 16E ALA E 187 SER E 190 TYPE III 1CSE 153 TURN 17 17E GLY E 193 LEU E 196 TYPE I 1CSE 154 TURN 18 18E GLY E 219 MET E 222 TYPE III 1CSE 155 TURN 19 19E HIS E 238 LEU E 241 TYPE I 1CSE 156 TURN 20 20E SER E 251 ALA E 254 TYPE I 1CSE 157 TURN 21 21E SER E 259 TYR E 262 TYPE III 1CSE 158 TURN 22 22E TYR E 263 GLY E 266 TYPE II(PRIME) 1CSE 159 TURN 23 23E ALA E 272 GLN E 275 TYPE I 1CSE 160 TURN 24 1I VAL I 13 LYS I 16 TYPE II 1CSE 161 TURN 25 2I TYR I 29 TYR I 32 TYPE I 1CSE 162 TURN 26 3I PRO I 38 SER I 41 TYPE II 1CSE 163 TURN 27 4I ARG I 48 ARG I 51 TYPE I 1CSE 164 TURN 28 5I ASN I 57 THR I 60 TYPE II 1CSE 165 SITE 1 ACT 3 ASP E 32 HIS E 64 SER E 221 1CSE 166 SITE 1 RSB 2 LEU I 45 ASP I 46 1CSE 167 CRYST1 38.300 41.500 57.000 111.80 85.80 104.70 P 1 1 1CSE 168 ORIGX1 1.000000 0.262345 0.024180 0.00000 1CSE 169 ORIGX2 0.000000 1.033840 0.406217 0.00000 1CSE 170 ORIGX3 0.000000 0.000000 1.077317 0.00000 1CSE 171 SCALE1 0.026110 0.006850 0.000631 0.00000 1CSE 172 SCALE2 0.000000 0.024912 0.009788 0.00000 1CSE 173 SCALE3 0.000000 0.000000 0.018900 0.00000 1CSE 174