HEADER HYDROLASE (SERINE PROTEINASE) 21-AUG-89 1S01 1S01 2 COMPND SUBTILISIN /BPN$(PRIME) 8350 (E.C.3.4.21.14) (MUTANT WITH 1S01 3 COMPND 2 MET 50 REPLACED BY PHE, ASN 76 REPLACED BY ASP, GLY 169 1S01 4 COMPND 3 REPLACED BY ALA, GLN 206 REPLACED BY CYS, TYR 217 REPLACED 1S01 5 COMPND 4 BY LYS, AND ASN 217 REPLACED BY SER) (/M50F$, /N76D$, 1S01 6 COMPND 5 /G169A$, /Q206C$, /Y217K$, AND /N218S$) IN /EDTA$ 1S01 7 SOURCE (BACILLUS $AMYLOLIQUEFACIENS) 1S01 8 AUTHOR M.WHITLOW,A.J.HOWARD,J.F.WOOD 1S01 9 REVDAT 1 15-OCT-90 1S01 0 1S01 10 JRNL AUTH M.W.PANTOLIANO,M.WHITLOW,J.F.WOOD,S.W.DODD, 1S01 11 JRNL AUTH 2 K.D.HARDMAN,M.L.ROLLENCE,,P.N.BRYAN 1S01 12 JRNL TITL LARGE INCREASES IN GENERAL STABILITY FOR 1S01 13 JRNL TITL 2 SUBTILISIN /BPN$(PRIME) THROUGH INCREMENTAL CHANGES 1S01 14 JRNL TITL 3 IN THE FREE ENERGY OF UNFOLDING 1S01 15 JRNL REF BIOCHEMISTRY V. 28 7205 1989 1S01 16 JRNL REFN ASTM BICHAW US ISSN 0006-2960 033 1S01 17 REMARK 1 1S01 18 REMARK 1 REFERENCE 1 1S01 19 REMARK 1 AUTH C.-*H.WONG,S.-*T.CHEN,W.J.HENNEN,J.A.BIBBS, 1S01 20 REMARK 1 AUTH 2 Y.-*F.WANG,J.L.-*C.LIU,M.W.PANTOLIANO,M.WHITLOW, 1S01 21 REMARK 1 AUTH 3 P.N.BRYAN 1S01 22 REMARK 1 TITL ENZYMES IN ORGANIC SYNTHESIS. USE OF SUBTILISIN AND 1S01 23 REMARK 1 TITL 2 A HIGHLY STABLE MUTANT DERIVED FROM MULTIPLE 1S01 24 REMARK 1 TITL 3 SITE-*SPECIFIC MUTATIONS 1S01 25 REMARK 1 REF J.AM.CHEM.SOC. V. 112 945 1990 1S01 26 REMARK 1 REFN ASTM JACSAT US ISSN 0002-7863 004 1S01 27 REMARK 1 REFERENCE 2 1S01 28 REMARK 1 AUTH M.W.PANTOLIANO,M.WHITLOW,J.F.WOOD,M.L.ROLLENCE, 1S01 29 REMARK 1 AUTH 2 B.C.FINZEL,G.L.GILLILAND,T.L.POULOS,P.N.BRYAN 1S01 30 REMARK 1 TITL THE ENGINEERING OF BINDING AFFINITY AT METAL ION 1S01 31 REMARK 1 TITL 2 BINDING SITES FOR THE STABILIZATION OF PROTEINS. 1S01 32 REMARK 1 TITL 3 SUBTILISIN AS A TEST CASE 1S01 33 REMARK 1 REF BIOCHEMISTRY V. 27 8311 1988 1S01 34 REMARK 1 REFN ASTM BICHAW US ISSN 0006-2960 033 1S01 35 REMARK 1 REFERENCE 3 1S01 36 REMARK 1 AUTH M.L.ROLLENCE,D.FILPULA,M.W.PANTOLIANO,P.N.BRYAN 1S01 37 REMARK 1 TITL ENGINEERING THERMOSTABILITY IN SUBTILISIN 1S01 38 REMARK 1 TITL 2 /BPN$(PRIME) BY IN VITRO MUTAGENESIS 1S01 39 REMARK 1 REF CRIT.REV.BIOTECHNOL. V. 8 217 1988 1S01 40 REMARK 1 REFN ASTM CRBTE5 US ISSN 0738-8551 819 1S01 41 REMARK 1 REFERENCE 4 1S01 42 REMARK 1 AUTH P.N.BRYAN,M.L.ROLLENCE,M.W.PANTOIANO,J.WOOD, 1S01 43 REMARK 1 AUTH 2 B.C.FINZEL,G.L.GILLILAND,A.J.HOWARD,T.L.POULOS 1S01 44 REMARK 1 TITL PROTEASES OF ENHANCED STABILITY. CHARACTERIZATION 1S01 45 REMARK 1 TITL 2 OF A THERMOSTABLE VARIANT OF SUBTILISIN 1S01 46 REMARK 1 REF PROTEINS.STRUCT.,FUNCT., V. 1 326 1986 1S01 47 REMARK 1 REF 2 GENET. 1S01 48 REMARK 1 REFN ASTM PSFGEY US ISSN 0887-3585 867 1S01 49 REMARK 1 REFERENCE 5 1S01 50 REMARK 1 AUTH R.A.ALDEN,J.J.BIRKTOFT,J.KRAUT,J.D.ROBERTUS, 1S01 51 REMARK 1 AUTH 2 C.S.WRIGHT 1S01 52 REMARK 1 TITL ATOMIC COORDINATES FOR SUBTILISIN /BPN$* (OR NOVO) 1S01 53 REMARK 1 REF BIOCHEM.BIOPHYS.RES.COMM. V. 45 337 1971 1S01 54 REMARK 1 REFN ASTM BBRCA9 US ISSN 0006-291X 146 1S01 55 REMARK 2 1S01 56 REMARK 2 RESOLUTION. 1.7 ANGSTROMS. 1S01 57 REMARK 3 1S01 58 REMARK 3 REFINEMENT. BY THE RESTRAINED LEAST-SQUARES PROCEDURE OF J. 1S01 59 REMARK 3 KONNERT AND W. HENDRICKSON AS MODIFIED BY B. FINZEL 1S01 60 REMARK 3 (PROGRAM *PROFFT*). THE R VALUE IS 0.152 FOR 17421 1S01 61 REMARK 3 REFLECTIONS IN THE RESOLUTION RANGE 10.0 TO 1.7 ANGSTROMS. 1S01 62 REMARK 3 1S01 63 REMARK 3 NUMBER OF PROTEIN ATOMS (INCLUDING CA, IPA) 1938 1S01 64 REMARK 3 NUMBER OF SOLVENT ATOMS 215 1S01 65 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES (THE VALUES OF 1S01 66 REMARK 3 SIGMA, IN PARENTHESES, ARE THE INPUT ESTIMATED 1S01 67 REMARK 3 STANDARD DEVIATIONS THAT DETERMINE THE RELATIVE 1S01 68 REMARK 3 WEIGHTS OF THE CORRESPONDING RESTRAINTS) 1S01 69 REMARK 3 DISTANCE RESTRAINTS (ANGSTROMS) 1S01 70 REMARK 3 BOND DISTANCE 0.017(0.030) 1S01 71 REMARK 3 ANGLE DISTANCE 0.027(0.040) 1S01 72 REMARK 3 PLANAR 1-4 DISTANCE 0.034(0.050) 1S01 73 REMARK 3 PLANE RESTRAINT (ANGSTROMS) 0.018(0.030) 1S01 74 REMARK 3 CHIRAL-CENTER RESTRAINT (ANGSTROMS**3) 0.189(0.300) 1S01 75 REMARK 3 NON-BONDED CONTACT RESTRAINTS (ANGSTROMS) 1S01 76 REMARK 3 SINGLE TORSION CONTACT 0.157(0.200) 1S01 77 REMARK 3 MULTIPLE TORSION CONTACT 0.133(0.200) 1S01 78 REMARK 3 POSSIBLE HYDROGEN BOND 0.154(0.200) 1S01 79 REMARK 3 CONFORMATIONAL TORSION ANGLE RESTRAINT (DEGREES) 1S01 80 REMARK 3 PLANAR (OMEGA) 3.1(7.5) 1S01 81 REMARK 3 STAGGERED 11.6(10.0) 1S01 82 REMARK 3 ORTHONORMAL 30.4(10.0) 1S01 83 REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS (ANGSTROMS**2) 1S01 84 REMARK 3 MAIN-CHAIN BOND .562(1.000) 1S01 85 REMARK 3 MAIN-CHAIN ANGLE .915(2.000) 1S01 86 REMARK 3 SIDE-CHAIN BOND 1.213(1.500) 1S01 87 REMARK 3 SIDE-CHAIN ANGLE 1.827(3.000) 1S01 88 REMARK 4 1S01 89 REMARK 4 RESIDUE 206 APPEARS TO BE CYSTEINE PERSULFIDE. ELECTRON 1S01 90 REMARK 4 DENSITY FOUND BEYOND THE PERSULFIDE HAS BEEN MODELLED AS A 1S01 91 REMARK 4 PARTIALLY OCCUPIED CARBON ATOM. THE ADDITIONAL SULFUR AND 1S01 92 REMARK 4 CARBON ARE PRESENTED ON *HETATM* RECORDS AS HET GROUP 1S01 93 REMARK 4 * CS*. THE EXACT CHEMICAL IDENTITY OF THIS GROUP IS, 1S01 94 REMARK 4 HOWEVER, UNKNOWN. SEE THE PAPER CITED ON THE *JRNL* 1S01 95 REMARK 4 RECORDS ABOVE. 1S01 96 SEQRES 1 275 ALA GLN SER VAL PRO TYR GLY VAL SER GLN ILE LYS ALA 1S01 97 SEQRES 2 275 PRO ALA LEU HIS SER GLN GLY TYR THR GLY SER ASN VAL 1S01 98 SEQRES 3 275 LYS VAL ALA VAL ILE ASP SER GLY ILE ASP SER SER HIS 1S01 99 SEQRES 4 275 PRO ASP LEU LYS VAL ALA GLY GLY ALA SER PHE VAL PRO 1S01 100 SEQRES 5 275 SER GLU THR PRO ASN PHE GLN ASP ASP ASN SER HIS GLY 1S01 101 SEQRES 6 275 THR HIS VAL ALA GLY THR VAL ALA ALA LEU ASP ASN SER 1S01 102 SEQRES 7 275 ILE GLY VAL LEU GLY VAL ALA PRO SER SER ALA LEU TYR 1S01 103 SEQRES 8 275 ALA VAL LYS VAL LEU GLY ASP ALA GLY SER GLY GLN TYR 1S01 104 SEQRES 9 275 SER TRP ILE ILE ASN GLY ILE GLU TRP ALA ILE ALA ASN 1S01 105 SEQRES 10 275 ASN MET ASP VAL ILE ASN MET SER LEU GLY GLY PRO SER 1S01 106 SEQRES 11 275 GLY SER ALA ALA LEU LYS ALA ALA VAL ASP LYS ALA VAL 1S01 107 SEQRES 12 275 ALA SER GLY VAL VAL VAL VAL ALA ALA ALA GLY ASN GLU 1S01 108 SEQRES 13 275 GLY SER THR GLY SER SER SER THR VAL GLY TYR PRO GLY 1S01 109 SEQRES 14 275 LYS TYR PRO SER VAL ILE ALA VAL GLY ALA VAL ASP ALA 1S01 110 SEQRES 15 275 SER ASN GLN ARG ALA SER PHE SER SER VAL GLY PRO GLU 1S01 111 SEQRES 16 275 LEU ASP VAL MET ALA PRO GLY VAL SER ILE CYS SER THR 1S01 112 SEQRES 17 275 LEU PRO GLY ASN LYS TYR GLY ALA LYS SER GLY THR SER 1S01 113 SEQRES 18 275 MET ALA SER PRO HIS VAL ALA GLY ALA ALA ALA LEU ILE 1S01 114 SEQRES 19 275 LEU SER LYS HIS PRO ASN TRP THR ASN THR GLN VAL ARG 1S01 115 SEQRES 20 275 SER SER LEU GLN ASN THR THR THR LYS LEU GLY ASP SER 1S01 116 SEQRES 21 275 PHE TYR TYR GLY LYS GLY LEU ILE ASN VAL GLN ALA ALA 1S01 117 SEQRES 22 275 ALA GLN 1S01 118 FTNOTE 1 1S01 119 FTNOTE 1 RESIDUE PRO 168 IS A CIS PROLINE. 1S01 120 FTNOTE 2 1S01 121 FTNOTE 2 RESIDUE 206 APPEARS TO BE CYSTEINE PERSULFIDE. ELECTRON 1S01 122 FTNOTE 2 DENSITY FOUND BEYOND THE PERSULFIDE HAS BEEN MODELLED AS A 1S01 123 FTNOTE 2 PARTIALLY OCCUPIED CARBON ATOM. THE ADDITIONAL SULFUR AND 1S01 124 FTNOTE 2 CARBON ARE PRESENTED ON *HETATM* RECORDS AS HET GROUP 1S01 125 FTNOTE 2 * CS*. THE EXACT CHEMICAL IDENTITY OF THIS GROUP IS, 1S01 126 FTNOTE 2 HOWEVER, UNKNOWN. SEE THE PAPER CITED ON THE *JRNL* 1S01 127 FTNOTE 2 RECORDS ABOVE. 1S01 128 HET CS 277 2 SEE FTNOTE 2. 1S01 129 HET IPA 290 4 ISOPROPYL ALCOHOL 1S01 130 HET IPA 291 4 ISOPROPYL ALCOHOL 1S01 131 HET CA 295 1 CALCIUM 1S01 132 FORMUL 2 CS C1 S1 1S01 133 FORMUL 3 IPA 2(C3 H8 O1) 1S01 134 FORMUL 4 CA CA1 1S01 135 FORMUL 5 HOH *215(H2 O1) 1S01 136 CRYST1 41.640 79.450 37.260 90.00 114.53 90.00 P 21 2 1S01 137 ORIGX1 1.000000 0.000000 0.000000 0.00000 1S01 138 ORIGX2 0.000000 1.000000 0.000000 0.00000 1S01 139 ORIGX3 0.000000 0.000000 1.000000 0.00000 1S01 140 SCALE1 0.024015 0.000000 0.009970 0.00000 1S01 141 SCALE2 0.000000 0.012587 0.000000 0.00000 1S01 142 SCALE3 0.000000 0.000000 0.029501 0.00000 1S01 143