HEADER PHOSPHATE TRANSPORT 20-JUL-94 1PBP 1PBP 2 COMPND TITLE: PHOSPHATE-BINDING PROTEIN (MUTANT WITH THR 141 1PBP 3 COMPND 2 REPLACED BY ASP) COMPLEXED WITH MONOBASIC PHOSPHATE ION. 1PBP 4 COMPND 3 MOLECULE: PHOSPHATE-BINDING PROTEIN 1PBP 5 COMPND 4 SUBSTITUTION_MUTATIONS: ( 1PBP 6 COMPND 5 NATIVE_RESIDUE: THR () 141() 1PBP 7 COMPND 6 MUTANT_RESIDUE: ASP () 141() 1PBP 8 COMPND 7 ) 1PBP 9 COMPND 8 HETEROGEN: MONOBASIC PHOSPHATE 1PBP 10 SOURCE (ESCHERICHIA COLI) 1PBP 11 AUTHOR Z.WANG,A.CHOUDHARY,P.S.LEDVINA,F.A.QUIOCHO 1PBP 12 REVDAT 1 15-OCT-94 1PBP 0 1PBP 13 JRNL AUTH Z.WANG,A.CHOUDHARY,P.S.LEDVINA,F.A.QUIOCHO 1PBP 14 JRNL TITL FINE TUNING OF THE SPECIFICITY OF THE PERIPLASMIC 1PBP 15 JRNL TITL 2 PHOSPHATE TRANSPORT RECEPTOR: SITE-DIRECTED 1PBP 16 JRNL TITL 3 MUTAGENESIS, LIGAND BINDING, AND CRYSTALLOGRAPHIC 1PBP 17 JRNL TITL 4 STUDIES 1PBP 18 JRNL REF TO BE PUBLISHED 1PBP 19 JRNL REFN 0353 1PBP 20 REMARK 1 1PBP 21 REMARK 1 REFERENCE 1 1PBP 22 REMARK 1 AUTH Z.CHANG,A.CHOUDHARY,R.LATHIGRA,F.A.QUIOCHO 1PBP 23 REMARK 1 TITL TH E IMMUNODOMINANT 38-KDA LIPOPROTEIN ANTIGEN OF 1PBP 24 REMARK 1 TITL 2 MYCOBACTERIUM TUBERCULOSIS IS A PHOSPHATE-BINDING 1PBP 25 REMARK 1 TITL 3 PROTEIN 1PBP 26 REMARK 1 REF J.BIOL.CHEM. V. 269 1956 1994 1PBP 27 REMARK 1 REFN ASTM JBCHA3 US ISSN 0021-9258 0071 1PBP 28 REMARK 1 REFERENCE 2 1PBP 29 REMARK 1 AUTH H.LUECKE,F.A.QUIOCHO 1PBP 30 REMARK 1 TITL HIGH SPECIFICITY OF A PHOSPHATE TRANSPORT PROTEIN 1PBP 31 REMARK 1 TITL 2 DETERMINED BY HYDROGEN BONDS 1PBP 32 REMARK 1 REF NATURE V. 347 402 1990 1PBP 33 REMARK 1 REFN ASTM NATUAS UK ISSN 0028-0836 0006 1PBP 34 REMARK 2 1PBP 35 REMARK 2 RESOLUTION. 1.90 ANGSTROMS. 1PBP 36 REMARK 3 1PBP 37 REMARK 3 REFINEMENT. 1PBP 38 REMARK 3 PROGRAM X-PLOR 1PBP 39 REMARK 3 AUTHORS BRUNGER 1PBP 40 REMARK 3 R VALUE 0.147 1PBP 41 REMARK 3 RMSD BOND DISTANCES 0.017 ANGSTROMS 1PBP 42 REMARK 3 RMSD BOND ANGLES 1.849 DEGREES 1PBP 43 REMARK 3 BOND DISTANCE WEIGHTS 0.06 ANGSTROMS 1PBP 44 REMARK 3 BOND ANGLE WEIGHTS 10.0 DEGREES 1PBP 45 REMARK 3 1PBP 46 REMARK 3 NUMBER OF REFLECTIONS 38401 1PBP 47 REMARK 3 RESOLUTION RANGE 8.0 - 1.9 ANGSTROMS 1PBP 48 REMARK 3 DATA CUTOFF 0. SIGMA(F) 1PBP 49 REMARK 3 COMPLETENESS FOR RANGE 69.5 PERCENT 1PBP 50 REMARK 3 1PBP 51 REMARK 3 DATA COLLECTION 1PBP 52 REMARK 3 NUMBER OF UNIQUE REFLECTIONS 22300 1PBP 53 REMARK 3 REJECTION CRITERIA 0. SIGMA(F) 1PBP 54 REMARK 3 COMPLETENESS OF DATA 69.5 PERCENT 1PBP 55 REMARK 3 1PBP 56 REMARK 3 NUMBER OF ATOMS PRESENT IN ENTRY 1PBP 57 REMARK 3 NUMBER OF PROTEIN ATOMS 2439 1PBP 58 REMARK 3 NUMBER OF SOLVENT ATOMS 214 1PBP 59 REMARK 3 NUMBER OF HETEROGEN ATOMS 5 1PBP 60 REMARK 4 1PBP 61 REMARK 4 THE LABORATORY HAS PREVIOUSLY DETERMINED THE 1.7 ANGSTROM 1PBP 62 REMARK 4 RESOLUTION STRUCTURE OF THE PHOSPHATE-BINDING PROTEIN, AN 1PBP 63 REMARK 4 INITIAL RECEPTOR FOR THE HIGH-AFFINITY ACTIVE TRANSPORT 1PBP 64 REMARK 4 SYSTEM OR PERMEASE IN E. COLI. IN THIS STUDY, THE AUTHORS 1PBP 65 REMARK 4 USE SITE-DIRECTED MUTAGENESIS TO GENERATE THE THR 141 TO 1PBP 66 REMARK 4 ASP MUTANT, AND SHOW THAT THIS MUTANT RESTRICTS BINDING OF 1PBP 67 REMARK 4 ONLY THE MONOBASIC PHOSPHATE, ABOLISHING BINDING OF THE 1PBP 68 REMARK 4 DIBASIC PHOSPHATE. 1PBP 69 REMARK 5 1PBP 70 REMARK 5 THE FOLLOWING SOLVENT MOLECULES LIE FARTHER THAN EXPECTED 1PBP 71 REMARK 5 FROM THE PROTEIN OR NUCLEIC ACID MOLECULE AND MAY BE 1PBP 72 REMARK 5 ASSOCIATED WITH A SYMMETRY-RELATED MOLECULE (M=MODEL 1PBP 73 REMARK 5 NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SEQ=SEQUENCE 1PBP 74 REMARK 5 NUMBER; I=INSERTION CODE): 1PBP 75 REMARK 5 1PBP 76 REMARK 5 M RES C SEQI 1PBP 77 REMARK 5 0 HOH 536 DISTANCE = 3.63 ANGSTROMS 1PBP 78 REMARK 5 0 HOH 537 DISTANCE = 5.13 ANGSTROMS 1PBP 79 REMARK 5 0 HOH 542 DISTANCE = 3.85 ANGSTROMS 1PBP 80 REMARK 5 0 HOH 569 DISTANCE = 3.53 ANGSTROMS 1PBP 81 REMARK 5 0 HOH 577 DISTANCE = 3.95 ANGSTROMS 1PBP 82 REMARK 5 0 HOH 594 DISTANCE = 3.70 ANGSTROMS 1PBP 83 REMARK 5 0 HOH 595 DISTANCE = 3.86 ANGSTROMS 1PBP 84 REMARK 5 0 HOH 603 DISTANCE = 4.14 ANGSTROMS 1PBP 85 REMARK 5 0 HOH 604 DISTANCE = 4.14 ANGSTROMS 1PBP 86 REMARK 5 0 HOH 611 DISTANCE = 3.54 ANGSTROMS 1PBP 87 REMARK 5 0 HOH 614 DISTANCE = 3.70 ANGSTROMS 1PBP 88 SEQRES 1 321 GLU ALA SER LEU THR GLY ALA GLY ALA THR PHE PRO ALA 1PBP 89 SEQRES 2 321 PRO VAL TYR ALA LYS TRP ALA ASP THR TYR GLN LYS GLU 1PBP 90 SEQRES 3 321 THR GLY ASN LYS VAL ASN TYR GLN GLY ILE GLY SER SER 1PBP 91 SEQRES 4 321 GLY GLY VAL LYS GLN ILE ILE ALA ASN THR VAL ASP PHE 1PBP 92 SEQRES 5 321 GLY ALA SER ASP ALA PRO LEU SER ASP GLU LYS LEU ALA 1PBP 93 SEQRES 6 321 GLN GLU GLY LEU PHE GLN PHE PRO THR VAL ILE GLY GLY 1PBP 94 SEQRES 7 321 VAL VAL LEU ALA VAL ASN ILE PRO GLY LEU LYS SER GLY 1PBP 95 SEQRES 8 321 GLU LEU VAL LEU ASP GLY LYS THR LEU GLY ASP ILE TYR 1PBP 96 SEQRES 9 321 LEU GLY LYS ILE LYS LYS TRP ASP ASP GLU ALA ILE ALA 1PBP 97 SEQRES 10 321 LYS LEU ASN PRO GLY LEU LYS LEU PRO SER GLN ASN ILE 1PBP 98 SEQRES 11 321 ALA VAL VAL ARG ARG ALA ASP GLY SER GLY ASP SER PHE 1PBP 99 SEQRES 12 321 VAL PHE THR SER TYR LEU ALA LYS VAL ASN GLU GLU TRP 1PBP 100 SEQRES 13 321 LYS ASN ASN VAL GLY THR GLY SER THR VAL LYS TRP PRO 1PBP 101 SEQRES 14 321 ILE GLY LEU GLY GLY LYS GLY ASN ASP GLY ILE ALA ALA 1PBP 102 SEQRES 15 321 PHE VAL GLN ARG LEU PRO GLY ALA ILE GLY TYR VAL GLU 1PBP 103 SEQRES 16 321 TYR ALA TYR ALA LYS GLN ASN ASN LEU ALA TYR THR LYS 1PBP 104 SEQRES 17 321 LEU ILE SER ALA ASP GLY LYS PRO VAL SER PRO THR GLU 1PBP 105 SEQRES 18 321 GLU ASN PHE ALA ASN ALA ALA LYS GLY ALA ASP TRP SER 1PBP 106 SEQRES 19 321 LYS THR PHE ALA GLN ASP LEU THR ASN GLN LYS GLY GLU 1PBP 107 SEQRES 20 321 ASP ALA TRP PRO ILE THR SER THR THR PHE ILE LEU ILE 1PBP 108 SEQRES 21 321 HIS LYS ASP GLN LYS LYS PRO GLU GLN GLY THR GLU VAL 1PBP 109 SEQRES 22 321 LEU LYS PHE PHE ASP TRP ALA TYR LYS THR GLY ALA LYS 1PBP 110 SEQRES 23 321 GLN ALA ASN ASP LEU ASP TYR ALA SER LEU PRO ASP SER 1PBP 111 SEQRES 24 321 VAL VAL GLU GLN VAL ARG ALA ALA TRP LYS THR ASN ILE 1PBP 112 SEQRES 25 321 LYS ASP SER SER GLY LYS PRO LEU TYR 1PBP 113 HET PO4 322 5 PHOSPHATE GROUP 1PBP 114 FORMUL 2 PO4 O4 P1 1PBP 115 FORMUL 3 HOH *214(H2 O1) 1PBP 116 CRYST1 41.880 64.570 124.670 90.00 90.00 90.00 P 21 21 21 4 1PBP 117 ORIGX1 1.000000 0.000000 0.000000 0.00000 1PBP 118 ORIGX2 0.000000 1.000000 0.000000 0.00000 1PBP 119 ORIGX3 0.000000 0.000000 1.000000 0.00000 1PBP 120 SCALE1 0.023878 0.000000 0.000000 0.00000 1PBP 121 SCALE2 0.000000 0.015487 0.000000 0.00000 1PBP 122 SCALE3 0.000000 0.000000 0.008021 0.00000 1PBP 123