HEADER IRON STORAGE 20-DEC-90 1FHA 1FHA 2 COMPND FERRITIN (H-CHAIN) MUTANT (LYS 86 REPLACED BY GLN) (K86Q) 1FHA 3 SOURCE HUMAN (HOMO $SAPIENS) RECOMBINANT FORM EXPRESSED IN 1FHA 4 SOURCE 2 (ESCHERICHIA $COLI) 1FHA 5 AUTHOR P.J.ARTYMIUK,P.M.HARRISON 1FHA 6 REVDAT 1 15-JUL-92 1FHA 0 1FHA 7 REMARK 1 1FHA 8 REMARK 1 REFERENCE 1 1FHA 9 REMARK 1 AUTH D.M.LAWSON,P.J.ARTYMIUK,S.J.YEWDALL,J.M.A.SMITH, 1FHA 10 REMARK 1 AUTH 2 J.C.LIVINGSTONE,A.TREFFRY,A.LUZZAGO,S.LEVI, 1FHA 11 REMARK 1 AUTH 3 P.AROSIO,G.CESARINI,C.D.THOMAS,W.V.SHAW, 1FHA 12 REMARK 1 AUTH 4 P.M.HARRISON 1FHA 13 REMARK 1 TITL SOLVING THE STRUCTURE OF HUMAN H FERRITIN BY 1FHA 14 REMARK 1 TITL 2 GENETICALLY ENGINEERING INTERMOLECULAR CRYSTAL 1FHA 15 REMARK 1 TITL 3 CONTACTS 1FHA 16 REMARK 1 REF NATURE V. 349 541 1991 1FHA 17 REMARK 1 REFN ASTM NATUAS UK ISSN 0028-0836 006 1FHA 18 REMARK 2 1FHA 19 REMARK 2 RESOLUTION. 2.4 ANGSTROMS. 1FHA 20 REMARK 3 1FHA 21 REMARK 3 REFINEMENT. BY THE RESTRAINED LEAST-SQUARES PROCEDURE OF J. 1FHA 22 REMARK 3 KONNERT AND W. HENDRICKSON (PROGRAM *PROLSQ*). THE R 1FHA 23 REMARK 3 VALUE IS 0.205. 1FHA 24 REMARK 3 1FHA 25 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES (THE VALUES OF 1FHA 26 REMARK 3 SIGMA, IN PARENTHESES, ARE THE INPUT ESTIMATED 1FHA 27 REMARK 3 STANDARD DEVIATIONS THAT DETERMINE THE RELATIVE 1FHA 28 REMARK 3 WEIGHTS OF THE CORRESPONDING RESTRAINTS) 1FHA 29 REMARK 3 DISTANCE RESTRAINTS (ANGSTROMS) 1FHA 30 REMARK 3 BOND DISTANCE 0.020(0.020) 1FHA 31 REMARK 3 ANGLE DISTANCE 0.036(0.025) 1FHA 32 REMARK 3 PLANAR 1-4 DISTANCE 0.047(0.025) 1FHA 33 REMARK 3 PLANE RESTRAINT (ANGSTROMS) 0.011(0.020) 1FHA 34 REMARK 3 CHIRAL-CENTER RESTRAINT (ANGSTROMS**3) 0.038(0.200) 1FHA 35 REMARK 3 NON-BONDED CONTACT RESTRAINTS (ANGSTROMS) 1FHA 36 REMARK 3 SINGLE TORSION CONTACT 0.210(0.500) 1FHA 37 REMARK 3 MULTIPLE TORSION CONTACT 0.196(0.500) 1FHA 38 REMARK 3 POSSIBLE HYDROGEN BOND 0.136(0.500) 1FHA 39 REMARK 4 1FHA 40 REMARK 4 THE ENTIRE FERRITIN MOLECULE FORMS A HOLLOW, APPROXIMATELY 1FHA 41 REMARK 4 SPHERICAL, ASSEMBLY CONSISTING OF 24 SUBUNITS RELATED BY 1FHA 42 REMARK 4 432 SYMMETRY. A COMPLETE 24-MER (CENTER AT X=0, Y=0, Z=0) 1FHA 43 REMARK 4 CAN BE GENERATED BY APPLYING THE 24 NON-TRANSLATIONAL 1FHA 44 REMARK 4 CRYSTALLOGRAPHIC OPERATIONS OF SPACE GROUP F 4 3 2. THESE 1FHA 45 REMARK 4 ARE: 1FHA 46 REMARK 4 1 X,Y,Z 1FHA 47 REMARK 4 2 X,-Y,-Z 1FHA 48 REMARK 4 3 -X,Y,-Z 1FHA 49 REMARK 4 4 -X,-Y,Z 1FHA 50 REMARK 4 5 Z,X,Y 1FHA 51 REMARK 4 6 Z,-X,-Y 1FHA 52 REMARK 4 7 -Z,X,-Y 1FHA 53 REMARK 4 8 -Z,-X,Y 1FHA 54 REMARK 4 9 Y,Z,X 1FHA 55 REMARK 4 10 Y,-Z,-X 1FHA 56 REMARK 4 11 -Y,Z,-X 1FHA 57 REMARK 4 12 -Y,-Z,X 1FHA 58 REMARK 4 13 -X,-Z,-Y 1FHA 59 REMARK 4 14 -X,Z,Y 1FHA 60 REMARK 4 15 X,-Z,Y 1FHA 61 REMARK 4 16 X,Z,-Y 1FHA 62 REMARK 4 17 -Y,-X,-Z 1FHA 63 REMARK 4 18 -Y,X,Z 1FHA 64 REMARK 4 19 Y,-X,Z 1FHA 65 REMARK 4 20 Y,X,-Z 1FHA 66 REMARK 4 21 -Z,-Y,-X 1FHA 67 REMARK 4 22 -Z,Y,X 1FHA 68 REMARK 4 23 Z,-Y,X 1FHA 69 REMARK 4 24 Z,Y,-X 1FHA 70 REMARK 5 1FHA 71 REMARK 5 THE STRUCTURE WAS SOLVED BY GENETICALLY ENGINEERING 1FHA 72 REMARK 5 INTERMOLECULAR CRYSTAL CONTACTS FROM THE KNOWN STRUCTURE OF 1FHA 73 REMARK 5 HORSE L CHAIN FERRITIN INTO HUMAN H CHAIN FERRITIN. 1FHA 74 REMARK 6 1FHA 75 REMARK 6 THE FERRITIN SEQUENCE CLONED IN ESCHERICHIA COLI WAS THAT 1FHA 76 REMARK 6 OF HOMO SAPIENS EXCEPT THAT LYS 86 WAS CHANGED TO GLN FOR 1FHA 77 REMARK 6 THE REASON GIVEN IN REMARK 5. 1FHA 78 REMARK 7 1FHA 79 REMARK 7 RESIDUES 1 - 4 WERE NOT VISIBLE IN THE ELECTRON DENSITY 1FHA 80 REMARK 7 MAP. SOME SIDE CHAINS ARE INCOMPLETE WHERE NO DENSITY WAS 1FHA 81 REMARK 7 VISIBLE. 1FHA 82 REMARK 8 1FHA 83 REMARK 8 RESIDUE NUMBERING WAS CHOSEN TO BE CONSISTENT WITH THE RAT 1FHA 84 REMARK 8 L CHAIN FERRITIN WHICH HAS AN EIGHT RESIDUE INSERTION 1FHA 85 REMARK 8 (RELATIVE TO HUMAN FERRITIN) FROM 162 - 169 (INCLUSIVE). 1FHA 86 REMARK 8 THESE EIGHT RESIDUES ARE NOT PRESENT IN HUMAN H FERRITIN. 1FHA 87 REMARK 8 THUS IN HUMAN H FERRITIN THE RESIDUE NUMBERING GOES 1FHA 88 REMARK 8 DIRECTLY FROM 161 TO 170 BUT NO RESIDUES ARE MISSING. 1FHA 89 REMARK 9 1FHA 90 REMARK 9 THE FERROXIDASE CENTER WHICH CATALYSES THE OXIDATION OF 1FHA 91 REMARK 9 FE(II) TO FE(III) INVOLVES THE FOLLOWING RESIDUES: 1FHA 92 REMARK 9 GLU 26, GLU 62, HIS 64, FE 200. COORDINATION 1FHA 93 REMARK 9 CONSIDERATIONS SUGGEST THAT THIS FE IS REALLY A CALCIUM 1FHA 94 REMARK 9 ION IN THIS STRUCTURE. 1FHA 95 REMARK 10 1FHA 96 REMARK 10 THERE IS A STRAND FROM 84 TO 86 THAT FORMS A TENUOUS 1FHA 97 REMARK 10 TWO-STRANDED ANIT-PARALLEL BETA SHEET WITH THE EQUIVALENT 1FHA 98 REMARK 10 STRAND IN A TWO-FOLD RELATED SUBUNIT. 1FHA 99 SEQRES 1 183 MET THR THR ALA SER THR SER GLN VAL ARG GLN ASN TYR 1FHA 100 SEQRES 2 183 HIS GLN ASP SER GLU ALA ALA ILE ASN ARG GLN ILE ASN 1FHA 101 SEQRES 3 183 LEU GLU LEU TYR ALA SER TYR VAL TYR LEU SER MET SER 1FHA 102 SEQRES 4 183 TYR TYR PHE ASP ARG ASP ASP VAL ALA LEU LYS ASN PHE 1FHA 103 SEQRES 5 183 ALA LYS TYR PHE LEU HIS GLN SER HIS GLU GLU ARG GLU 1FHA 104 SEQRES 6 183 HIS ALA GLU LYS LEU MET LYS LEU GLN ASN GLN ARG GLY 1FHA 105 SEQRES 7 183 GLY ARG ILE PHE LEU GLN ASP ILE GLN LYS PRO ASP CYS 1FHA 106 SEQRES 8 183 ASP ASP TRP GLU SER GLY LEU ASN ALA MET GLU CYS ALA 1FHA 107 SEQRES 9 183 LEU HIS LEU GLU LYS ASN VAL ASN GLN SER LEU LEU GLU 1FHA 108 SEQRES 10 183 LEU HIS LYS LEU ALA THR ASP LYS ASN ASP PRO HIS LEU 1FHA 109 SEQRES 11 183 CYS ASP PHE ILE GLU THR HIS TYR LEU ASN GLU GLN VAL 1FHA 110 SEQRES 12 183 LYS ALA ILE LYS GLU LEU GLY ASP HIS VAL THR ASN LEU 1FHA 111 SEQRES 13 183 ARG LYS MET GLY ALA PRO GLU SER GLY LEU ALA GLU TYR 1FHA 112 SEQRES 14 183 LEU PHE ASP LYS HIS THR LEU GLY ASP SER ASP ASN GLU 1FHA 113 SEQRES 15 183 SER 1FHA 114 FTNOTE 1 1FHA 115 FTNOTE 1 RESIDUE 161 IS A CIS PROLINE. 1FHA 116 CRYST1 184.800 184.800 184.800 90.00 90.00 90.00 F 4 3 2 24 1FHA 117 ORIGX1 1.000000 0.000000 0.000000 0.00000 1FHA 118 ORIGX2 0.000000 1.000000 0.000000 0.00000 1FHA 119 ORIGX3 0.000000 0.000000 1.000000 0.00000 1FHA 120 SCALE1 0.005411 0.000000 0.000000 0.00000 1FHA 121 SCALE2 0.000000 0.005411 0.000000 0.00000 1FHA 122 SCALE3 0.000000 0.000000 0.005411 0.00000 1FHA 123 HET FE 200 1 IRON 1FHA 124 HET CA 210 1 CALCIUM 1FHA 125 HET CA 211 1 CALCIUM 1FHA 126 FORMUL 2 FE FE1 1FHA 127 FORMUL 3 CA 2(CA1) 1FHA 128 FORMUL 4 HOH *20(H2 O1) 1FHA 129 HELIX 1 A GLN 14 ARG 43 1 1FHA 130 HELIX 2 B ASN 50 ARG 76 1 1FHA 131 HELIX 3 C GLY 96 ASP 123 1 1FHA 132 HELIX 4 D LEU 129 MET 158 1 KINKED AT RESIDUES 136-137 1FHA 133 HELIX 5 E SER 171 LYS 180 1 1FHA 134