USER MOD reduce.3.24.130724 H: found=0, std=0, add=14, rem=0, adj=0 USER MOD reduce.3.24.130724 removed 14 hydrogens (0 hets) HEADER ACETYLCHOLINE RECEPTOR 09-MAR-99 1CEK TITLE THREE-DIMENSIONAL STRUCTURE OF THE MEMBRANE-EMBEDDED M2 TITLE 2 CHANNEL-LINING SEGMENT FROM THE NICOTINIC ACETYLCHOLINE TITLE 3 RECEPTOR BY SOLID-STATE NMR SPECTROSCOPY COMPND MOL_ID: 1; COMPND 2 MOLECULE: PROTEIN (ACETYLCHOLINE RECEPTOR M2); COMPND 3 CHAIN: A; COMPND 4 FRAGMENT: M2 DOMAIN; COMPND 5 SYNONYM: ACHR M2; COMPND 6 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: RATTUS NORVEGICUS; SOURCE 3 ORGANISM_COMMON: NORWAY RAT; SOURCE 4 ORGANISM_TAXID: 10116; SOURCE 5 ORGAN: BRAIN; SOURCE 6 CELL: NEURON; SOURCE 7 CELLULAR_LOCATION: POST-SYNAPTIC MEMBRANE; SOURCE 8 EXPRESSION_SYSTEM: ESCHERICHIA COLI BL21; SOURCE 9 EXPRESSION_SYSTEM_TAXID: 511693; SOURCE 0 EXPRESSION_SYSTEM_STRAIN: BL21; SOURCE 1 EXPRESSION_SYSTEM_VECTOR_TYPE: BACTERIAL; SOURCE 2 EXPRESSION_SYSTEM_PLASMID: PGEX FUSION KEYWDS ACETYLCHOLINE RECEPTOR, M2, LIPID BILAYERS, ION-CHANNEL EXPDTA SOLID-STATE NMR AUTHOR F.M.MARASSI,J.J.GESELL,Y.KIM,A.P.VALENTE,M.OBLATT-MONTAL, AUTHOR 2 M.MONTAL,S.J.OPELLA REVDAT 3 24-FEB-09 1CEK 1 VERSN REVDAT 2 09-APR-99 1CEK 1 JRNL REVDAT 1 11-MAR-99 1CEK 0 JRNL AUTH S.J.OPELLA,F.M.MARASSI,J.J.GESELL,A.P.VALENTE, JRNL AUTH 2 Y.KIM,M.OBLATT-MONTAL,M.MONTAL JRNL TITL STRUCTURES OF THE M2 CHANNEL-LINING SEGMENTS FROM JRNL TITL 2 NICOTINIC ACETYLCHOLINE AND NMDA RECEPTORS BY NMR JRNL TITL 3 SPECTROSCOPY. JRNL REF NAT.STRUCT.BIOL. V. 6 374 1999 JRNL REFN ISSN 1072-8368 JRNL PMID 10201407 JRNL DOI 10.1038/7610 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH F.M.MARASSI,J.J.GESELL,A.P.VALENTE,Y.KIM, REMARK 1 AUTH 2 M.OBLATT-MONTAL,M.MONTAL,S.J.OPELLA REMARK 1 TITL DILUTE SPIN-EXCHANGE ASSIGNMENT OF SOLID-STATE NMR REMARK 1 TITL 2 SPECTRA OF ORIENTED PROTEINS: ACETYLCHOLINE REMARK 1 TITL 3 RECEPTOR M2 PEPTIDE IN BILAYERS REMARK 1 REF TO BE PUBLISHED REMARK 1 REFN REMARK 2 REMARK 2 RESOLUTION. NOT APPLICABLE. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : RESTRICT REMARK 3 AUTHORS : NULL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: NULL REMARK 4 REMARK 4 1CEK COMPLIES WITH FORMAT V. 3.15, 01-DEC-08 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 11-MAR-99. REMARK 100 THE RCSB ID CODE IS RCSB000606. REMARK 210 REMARK 210 EXPERIMENTAL DETAILS REMARK 210 EXPERIMENT TYPE : NMR REMARK 210 TEMPERATURE (KELVIN) : 295 REMARK 210 PH : 6.0 REMARK 210 IONIC STRENGTH : NULL REMARK 210 PRESSURE : NULL REMARK 210 SAMPLE CONTENTS : DMPC REMARK 210 REMARK 210 NMR EXPERIMENTS CONDUCTED : 15N SHIFT/1H-15N DIPOLAR REMARK 210 COUPLING PISEMA, 15N SHIFT/15N REMARK 210 SHIFT HETCOR, 1H SHIFT/15N REMARK 210 SHIFT/1H-15N DIPOLAR COUPLING REMARK 210 CORRELATION REMARK 210 SPECTROMETER FIELD STRENGTH : 400 MHZ, 550 MHZ REMARK 210 SPECTROMETER MODEL : HOMEBUILT, CMX400 REMARK 210 SPECTROMETER MANUFACTURER : HOMEBUILT, CHEMAGNETICS REMARK 210 REMARK 210 STRUCTURE DETERMINATION. REMARK 210 SOFTWARE USED : FISI, BACKTOR, RESTRICT REMARK 210 METHOD USED : ORIENTATIONAL CONSTRAINTS REMARK 210 REMARK 210 CONFORMERS, NUMBER CALCULATED : 24 REMARK 210 CONFORMERS, NUMBER SUBMITTED : 1 REMARK 210 CONFORMERS, SELECTION CRITERIA : LOWEST ENERGY REMARK 210 REMARK 210 BEST REPRESENTATIVE CONFORMER IN THIS ENSEMBLE : NULL REMARK 210 REMARK 210 REMARK: LOWEST ENERGY. SAMPLE: DMPC BILAYERS ORIENTED ON GLASS REMARK 210 SLIDES 15N CHEMICAL SHIFT, 1H CHEMICAL SHIFT AND 1H-15N REMARK 210 DIPOLAR COUPLING FREQUENCIES WERE MEASURED FROM PISEMA AND REMARK 217 REMARK 217 SOLID STATE NMR STUDY REMARK 217 THE COORDINATES IN THIS ENTRY WERE GENERATED FROM SOLID REMARK 217 STATE NMR DATA. PROTEIN DATA BANK CONVENTIONS REQUIRE THAT REMARK 217 CRYST1 AND SCALE RECORDS BE INCLUDED, BUT THE VALUES ON REMARK 217 THESE RECORDS ARE MEANINGLESS. REMARK 465 REMARK 465 MISSING RESIDUES REMARK 465 THE FOLLOWING RESIDUES WERE NOT LOCATED IN THE REMARK 465 EXPERIMENT. (RES=RESIDUE NAME; C=CHAIN IDENTIFIER; REMARK 465 SSSEQ=SEQUENCE NUMBER; I=INSERTION CODE.) REMARK 465 RES C SSSEQI REMARK 465 GLY A 1 REMARK 465 SER A 2 REMARK 465 GLU A 3 REMARK 465 LYS A 4 REMARK 465 MET A 5 REMARK 465 SER A 6 REMARK 465 THR A 22 REMARK 465 SER A 23 REMARK 465 GLN A 24 REMARK 465 ARG A 25 REMARK 470 REMARK 470 MISSING ATOM REMARK 470 THE FOLLOWING RESIDUES HAVE MISSING ATOMS(RES=RESIDUE NAME; REMARK 470 C=CHAIN IDENTIFIER; SSEQ=SEQUENCE NUMBER; I=INSERTION CODE): REMARK 470 RES CSSEQI ATOMS REMARK 470 THR A 7 N CB OG1 CG2 REMARK 470 ALA A 8 CB REMARK 470 ILE A 9 CB CG1 CG2 CD1 REMARK 470 SER A 10 CB OG REMARK 470 VAL A 11 CB CG1 CG2 REMARK 470 LEU A 12 CB CG CD1 CD2 REMARK 470 LEU A 13 CB CG CD1 CD2 REMARK 470 ALA A 14 CB REMARK 470 GLN A 15 CB CG CD OE1 NE2 REMARK 470 ALA A 16 CB REMARK 470 VAL A 17 CB CG1 CG2 REMARK 470 PHE A 18 CB CG CD1 CD2 CE1 CE2 CZ REMARK 470 LEU A 19 CB CG CD1 CD2 REMARK 470 LEU A 20 CB CG CD1 CD2 REMARK 470 LEU A 21 C O CB CG CD1 CD2 DBREF 1CEK A 1 25 UNP P25110 ACHD_RAT 274 298 SEQADV 1CEK GLY A 1 UNP P25110 CYS 274 CLONING ARTIFACT SEQADV 1CEK SER A 2 UNP P25110 GLY 275 CLONING ARTIFACT SEQADV 1CEK MET A 5 UNP P25110 THR 278 CONFLICT SEQADV 1CEK THR A 7 UNP P25110 VAL 280 CONFLICT SEQADV 1CEK ALA A 16 UNP P25110 SER 289 CONFLICT SEQADV 1CEK THR A 22 UNP P25110 ILE 295 CONFLICT SEQADV 1CEK GLN A 24 UNP P25110 LYS 297 CONFLICT SEQRES 1 A 25 GLY SER GLU LYS MET SER THR ALA ILE SER VAL LEU LEU SEQRES 2 A 25 ALA GLN ALA VAL PHE LEU LEU LEU THR SER GLN ARG HELIX 1 1 THR A 7 LEU A 21 1SEE REMARK 650 15 CRYST1 1.000 1.000 1.000 90.00 90.00 90.00 P 1 1 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 1.000000 0.000000 0.000000 0.00000 SCALE2 0.000000 1.000000 0.000000 0.00000 SCALE3 0.000000 0.000000 1.000000 0.00000 ATOM 1 CA THR A 7 -0.360 0.941 -2.200 1.00 0.00 C ATOM 2 C THR A 7 0.085 -0.223 -1.315 1.00 0.00 C ATOM 3 O THR A 7 0.492 -1.284 -1.812 1.00 0.00 O ATOM 4 N ALA A 8 0.000 0.000 0.000 1.00 0.00 N ATOM 5 CA ALA A 8 0.389 -1.016 0.957 1.00 0.00 C ATOM 6 C ALA A 8 -0.592 -2.186 0.907 1.00 0.00 C ATOM 7 O ALA A 8 -0.344 -3.253 1.487 1.00 0.00 O ATOM 0 H ALA A 8 -0.333 0.871 0.414 1.00 0.00 H new ATOM 9 N ILE A 9 -1.707 -1.957 0.203 1.00 0.00 N ATOM 10 CA ILE A 9 -2.726 -2.977 0.071 1.00 0.00 C ATOM 11 C ILE A 9 -2.500 -3.779 -1.209 1.00 0.00 C ATOM 12 O ILE A 9 -2.971 -4.917 -1.341 1.00 0.00 O ATOM 0 H ILE A 9 -1.915 -1.081 -0.275 1.00 0.00 H new ATOM 14 N SER A 10 -1.769 -3.159 -2.142 1.00 0.00 N ATOM 15 CA SER A 10 -1.475 -3.800 -3.407 1.00 0.00 C ATOM 16 C SER A 10 -0.471 -4.934 -3.196 1.00 0.00 C ATOM 17 O SER A 10 -0.521 -5.966 -3.880 1.00 0.00 O ATOM 0 H SER A 10 -1.378 -2.223 -2.037 1.00 0.00 H new ATOM 19 N VAL A 11 0.436 -4.717 -2.238 1.00 0.00 N ATOM 20 CA VAL A 11 1.449 -5.706 -1.930 1.00 0.00 C ATOM 21 C VAL A 11 0.792 -6.983 -1.406 1.00 0.00 C ATOM 22 O VAL A 11 1.234 -8.097 -1.708 1.00 0.00 O ATOM 0 H VAL A 11 0.482 -3.870 -1.672 1.00 0.00 H new ATOM 24 N LEU A 12 -0.268 -6.793 -0.617 1.00 0.00 N ATOM 25 CA LEU A 12 -0.988 -7.915 -0.048 1.00 0.00 C ATOM 26 C LEU A 12 -1.426 -8.871 -1.157 1.00 0.00 C ATOM 27 O LEU A 12 -1.260 -10.040 -1.075 1.00 0.00 O ATOM 0 H LEU A 12 -0.638 -5.877 -0.364 1.00 0.00 H new ATOM 29 N LEU A 13 -1.985 -8.345 -2.190 1.00 0.00 N ATOM 30 CA LEU A 13 -2.448 -9.137 -3.314 1.00 0.00 C ATOM 31 C LEU A 13 -1.289 -9.948 -3.894 1.00 0.00 C ATOM 32 O LEU A 13 -1.480 -11.064 -4.398 1.00 0.00 O ATOM 0 H LEU A 13 -2.143 -7.343 -2.296 1.00 0.00 H new ATOM 34 N ALA A 14 -0.089 -9.359 -3.811 1.00 0.00 N ATOM 35 CA ALA A 14 1.099 -10.012 -4.321 1.00 0.00 C ATOM 36 C ALA A 14 1.590 -11.062 -3.322 1.00 0.00 C ATOM 37 O ALA A 14 2.231 -12.052 -3.699 1.00 0.00 O ATOM 0 H ALA A 14 0.073 -8.441 -3.398 1.00 0.00 H new ATOM 39 N GLN A 15 1.272 -10.818 -2.044 1.00 0.00 N ATOM 40 CA GLN A 15 1.671 -11.728 -0.988 1.00 0.00 C ATOM 41 C GLN A 15 0.813 -12.992 -1.034 1.00 0.00 C ATOM 42 O GLN A 15 1.327 -14.114 -1.007 1.00 0.00 O ATOM 0 H GLN A 15 0.745 -10.004 -1.729 1.00 0.00 H new ATOM 44 N ALA A 16 -0.502 -12.781 -1.104 1.00 0.00 N ATOM 45 CA ALA A 16 -1.437 -13.889 -1.154 1.00 0.00 C ATOM 46 C ALA A 16 -1.343 -14.591 -2.510 1.00 0.00 C ATOM 47 O ALA A 16 -1.864 -15.702 -2.690 1.00 0.00 O ATOM 0 H ALA A 16 -0.935 -11.858 -1.127 1.00 0.00 H new ATOM 49 N VAL A 17 -0.669 -13.918 -3.451 1.00 0.00 N ATOM 50 CA VAL A 17 -0.501 -14.465 -4.783 1.00 0.00 C ATOM 51 C VAL A 17 0.619 -15.504 -4.784 1.00 0.00 C ATOM 52 O VAL A 17 0.488 -16.586 -5.376 1.00 0.00 O ATOM 0 H VAL A 17 -0.239 -13.004 -3.307 1.00 0.00 H new ATOM 54 N PHE A 18 1.720 -15.149 -4.109 1.00 0.00 N ATOM 55 CA PHE A 18 2.864 -16.036 -4.026 1.00 0.00 C ATOM 56 C PHE A 18 2.470 -17.334 -3.319 1.00 0.00 C ATOM 57 O PHE A 18 2.741 -18.434 -3.806 1.00 0.00 O ATOM 0 H PHE A 18 1.833 -14.261 -3.620 1.00 0.00 H new ATOM 59 N LEU A 19 1.824 -17.176 -2.164 1.00 0.00 N ATOM 60 CA LEU A 19 1.390 -18.320 -1.386 1.00 0.00 C ATOM 61 C LEU A 19 0.495 -19.222 -2.237 1.00 0.00 C ATOM 62 O LEU A 19 0.551 -20.395 -2.184 1.00 0.00 O ATOM 0 H LEU A 19 1.594 -16.271 -1.754 1.00 0.00 H new ATOM 64 N LEU A 20 -0.326 -18.647 -3.018 1.00 0.00 N ATOM 65 CA LEU A 20 -1.231 -19.383 -3.879 1.00 0.00 C ATOM 66 C LEU A 20 -0.446 -20.069 -4.998 1.00 0.00 C ATOM 67 O LEU A 20 -0.758 -21.201 -5.396 1.00 0.00 O ATOM 0 H LEU A 20 -0.410 -17.634 -3.098 1.00 0.00 H new ATOM 69 N LEU A 21 0.575 -19.356 -5.492 1.00 0.00 N ATOM 70 CA LEU A 21 1.405 -19.882 -6.557 1.00 0.00 C ATOM 0 H LEU A 21 0.835 -18.424 -5.168 1.00 0.00 H new TER 72 LEU A 21 END