HEADER HYDROLASE(O-GLYCOSYL) 28-MAY-93 119L COMPND LYSOZYME (E.C.3.2.1.17) MUTANT WITH CYS 54 REPLACED BY THR, COMPND 2 CYS 97 REPLACED BY ALA, ALA 134 REPLACED BY SER (C54T,C97A, COMPND 3 A134S) SOURCE BACTERIOPHAGE T4 (MUTANT GENE DERIVED FROM THE M13 SOURCE 2 PLASMID BY CLONING THE T4 LYSOZYME GENE) AUTHOR M.BLABER,B.W.MATTHEWS REVDAT 1 31-OCT-93 119L 0 JRNL AUTH M.BLABER,J.D.LINDSTROM,N.GASSNER,J.XU,D.W.HEINZ, JRNL AUTH 2 B.W.MATTHEWS JRNL TITL THE ENERGETIC COST AND THE STRUCTURAL JRNL TITL 2 CONSEQUENCES OF BURYING A HYDROXYL GROUP WITHIN JRNL TITL 3 THE CORE OF A PROTEIN DETERMINED FROM ALA TO SER JRNL TITL 4 AND VAL TO THR SUBSTITUTIONS IN T4 LYSOZYME JRNL REF TO BE PUBLISHED JRNL REFN 353 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH M.MATSUMURA,B.W.MATTHEWS REMARK 1 TITL CONTROL OF ENZYME ACTIVITY BY AN ENGINEERED REMARK 1 TITL 2 DISULFIDE BOND REMARK 1 REF SCIENCE V. 243 792 1989 REMARK 1 REFN ASTM SCIEAS US ISSN 0036-8075 038 REMARK 1 REFERENCE 2 REMARK 1 AUTH L.H.WEAVER,B.W.MATTHEWS REMARK 1 TITL STRUCTURE OF BACTERIOPHAGE T4 LYSOZYME REFINED AT REMARK 1 TITL 2 1.7 ANGSTROMS RESOLUTION REMARK 1 REF J.MOL.BIOL. V. 193 189 1987 REMARK 1 REFN ASTM JMOBAK UK ISSN 0022-2836 070 REMARK 2 REMARK 2 RESOLUTION. 1.65 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM TNT REMARK 3 AUTHORS TRONRUD,TEN EYCK,MATTHEWS REMARK 3 R VALUE 0.162 REMARK 3 RMSD BOND DISTANCES 0.017 ANGSTROMS REMARK 3 RMSD BOND ANGLES 2.0 DEGREES REMARK 4 REMARK 4 THERE ARE A LARGE NUMBER OF PROTEIN DATA BANK ENTRIES THAT REMARK 4 PRESENT COORDINATES FOR MOLECULES PRODUCED BY SITE-DIRECTED REMARK 4 MUTAGENESIS OF BACTERIOPHAGE T4 LYSOZYME. COORDINATES OF REMARK 4 THE NATIVE ENZYME ARE PRESENTED IN PROTEIN DATA BANK REMARK 4 ENTRIES 2LZM (DESIGNATED NATR1 BY THE DEPOSITORS) AND 3LZM REMARK 4 (DESIGNATED T4167RF BY THE DEPOSITORS). 3LZM SUPERSEDES REMARK 4 2LZM AND IS BASED ON ADDITIONAL REFINEMENT USING REMARK 4 RECOLLECTED DATA TO 1.7 ANGSTROMS RESOLUTION. HOWEVER, REMARK 4 BOTH 3LZM AND 2LZM ARE AVAILABLE FROM THE PROTEIN DATA REMARK 4 BANK. COMPARISON OF A MUTANT LYSOZYME TO WILD-TYPE REMARK 4 LYSOZYME IS BEST DONE BY USING THE WILD-TYPE COORDINATE SET REMARK 4 (EITHER 3LZM OR 2LZM) WHICH WAS USED AS THE STARTING POINT REMARK 4 FOR THE REFINEMENT OF THAT MUTANT. REMARK 4 REMARK 4 THIS STRUCTURE IS ISOMORPHOUS TO WILD TYPE. REMARK 4 STARTING COORDINATES WERE BASED ON THE WILD-TYPE MODEL. REMARK 5 REMARK 5 THE ORTHOGONAL X,Y,Z AXES OF THIS COORDINATE SET ARE REMARK 5 ALIGNED IN THE A*,B,C CRYSTALLOGRAPHIC DIRECTIONS. REMARK 6 REMARK 6 RESIDUES 162 - 164 IN WILD-TYPE AND ALL MUTANT LYSOZYMES REMARK 6 ARE EXTREMELY MOBILE. THUS THE COORDINATES FOR THESE REMARK 6 RESIDUES ARE VERY UNRELIABLE. THIS ENTRY DOES NOT INCLUDE REMARK 6 RESIDUES 163 AND 164. REMARK 7 REMARK 7 THERE ARE SEVERAL SUBTLE ASPECTS OF THE SECONDARY STRUCTURE REMARK 7 OF THIS MOLECULE WHICH CANNOT CONVENIENTLY BE REPRESENTED REMARK 7 IN THE HELIX AND SHEET RECORDS BELOW. THESE ASPECTS REMARK 7 INFLUENCE THE REPRESENTATION OF HELIX 6 AND STRAND 3 OF REMARK 7 SHEET *S1*. THE PAPER J.MOL.BIOL., V. 118, P. 81, 1978 REMARK 7 SHOULD BE CONSULTED FOR THESE SUBTLETIES. REMARK 8 REMARK 8 SEO 901 FORMS AN S-S LINKAGE TO SEO 902. THE DEPOSITORS REMARK 8 ASSIGNED THE RESIDUE NAME BME TO THE BETA-MERCAPTOETHANOL. REMARK 8 IN ORDER TO FOLLOW PROTEIN DATA BANK NOMENCLATURE, THE REMARK 8 THE MERCAPTOETHANOLS ARE PRESENTED AS *HET* GROUPS *SEO*. SEQRES 1 164 MET ASN ILE PHE GLU MET LEU ARG ILE ASP GLU GLY LEU SEQRES 2 164 ARG LEU LYS ILE TYR LYS ASP THR GLU GLY TYR TYR THR SEQRES 3 164 ILE GLY ILE GLY HIS LEU LEU THR LYS SER PRO SER LEU SEQRES 4 164 ASN ALA ALA LYS SER GLU LEU ASP LYS ALA ILE GLY ARG SEQRES 5 164 ASN THR ASN GLY VAL ILE THR LYS ASP GLU ALA GLU LYS SEQRES 6 164 LEU PHE ASN GLN ASP VAL ASP ALA ALA VAL ARG GLY ILE SEQRES 7 164 LEU ARG ASN ALA LYS LEU LYS PRO VAL TYR ASP SER LEU SEQRES 8 164 ASP ALA VAL ARG ARG ALA ALA LEU ILE ASN MET VAL PHE SEQRES 9 164 GLN MET GLY GLU THR GLY VAL ALA GLY PHE THR ASN SER SEQRES 10 164 LEU ARG MET LEU GLN GLN LYS ARG TRP ASP GLU ALA ALA SEQRES 11 164 VAL ASN LEU SER LYS SER ARG TRP TYR ASN GLN THR PRO SEQRES 12 164 ASN ARG ALA LYS ARG VAL ILE THR THR PHE ARG THR GLY SEQRES 13 164 THR TRP ASP ALA TYR LYS ASN LEU FTNOTE 1 FTNOTE 1 RESIDUES 162 - 164 IN WILD-TYPE AND ALL MUTANT LYSOZYMES FTNOTE 1 ARE EXTREMELY MOBILE. THUS THE COORDINATES FOR THESE FTNOTE 1 RESIDUES ARE VERY UNRELIABLE. THIS ENTRY DOES NOT INCLUDE FTNOTE 1 RESIDUES 163 AND 164. FTNOTE 2 FTNOTE 2 SEO 901 FORMS AN S-S LINKAGE TO SEO 902. HET SEO 901 4 BETA-MERCAPTOETHANOL HET SEO 902 4 BETA-MERCAPTOETHANOL HET CL 173 1 CHLORIDE ION HET CL 178 1 CHLORIDE ION FORMUL 2 SEO 2(C2 H6 O1 S1) FORMUL 3 CL 2(CL1 -) FORMUL 4 HOH *161(H2 O1) HELIX 1 H1 ILE 3 GLU 11 1 HELIX 2 H2 LEU 39 ILE 50 1 HELIX 3 H3 LYS 60 ARG 80 1 HELIX 4 H4 ALA 82 SER 90 1 HELIX 5 H5 ALA 93 MET 106 1 HELIX 6 H6 GLU 108 GLY 113 1 HELIX 7 H7 THR 115 GLN 123 1 HELIX 8 H8 TRP 126 SER 134 1 HELIX 9 H9 ARG 137 GLN 141 1 HELIX 10 H10 PRO 143 THR 155 1 TURN 1 T1 ASP 20 GLY 23 TURN 2 T2 THR 54 VAL 57 CRYST1 61.300 61.300 97.300 90.00 90.00 120.00 P 32 2 1 6 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 0.016313 0.009418 0.000000 0.00000 SCALE2 0.000000 0.018837 0.000000 0.00000 SCALE3 0.000000 0.000000 0.010277 0.00000