data_17098 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Cofactor fingerprinting with STD NMR to characterize proteins of unknown function: identification of a rare cCMP cofactor preference ; _BMRB_accession_number 17098 _BMRB_flat_file_name bmr17098.str _Entry_type original _Submission_date 2010-08-02 _Accession_date 2010-08-02 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details . loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Yao Huili . . 2 Sem Daniel S. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count chemical_rates 1 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2010-11-10 original author . stop_ _Original_release_date 2010-11-10 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'Cofactor fingerprinting with STD NMR to characterize proteins of unknown function: identification of a rare cCMP cofactor preference' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 15670825 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Yao Huili . . 2 Sem Daniel S. . stop_ _Journal_abbreviation 'FEBS Lett.' _Journal_name_full 'FEBS Letters' _Journal_volume 579 _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 661 _Page_last 666 _Year 2005 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'RSP2/ATP complex' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label RSP2 $RSP2 ATP $ATP stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_RSP2 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common RSP2 _Molecular_mass . _Mol_thiol_state 'not reported' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 738 _Mol_residue_sequence ; MAPTQAGHDTAYLKETVGEA LARGCAAAISAQPNDPVEYL GLWLLKYVKNAEVEGNFYRE RQQDLQKKKDRLVKEAQSEQ AAKSVALTRKEAADALALVT AEPRELLEAAVKLVKQHTAA GAAYAAVVAEPEEPDWVAPE DDEAAAVETEDEAAGGAALA EGEEPPPEPEPEPEAAPEDG EGDAPAPKIPRPVDYSKKYF AYVAASAGQEHVLEADLYRP APPPEDADEDFKPEPLPYSF RVLDEKLPMLYVPNVAAEER VKFFRKFPKIGSYQACGVAL PASGEFKALLAADTLFPEGS GQPLSADDRDFVWEVSQSLS RALEAVQARAAEALAATSAA EAVEELKAKVAELREQAAAE AAAAAPPPPAEGEEGEGEAP PAEEEPPAEEEAEEEEEEAE EGAEEGAEEGEEGEEAPPKP KKKKKVFNPIPGLQAAIEKL TAAAEAATEADARAQAAVAL EKQALDEVVALASSHSDATL SSLRNMLSVPQGTYHVVKAL LHLLGRPAASFSTWKRAHSH FSPRLFEDMAAYDATAERDM AVWGRVRSCYKAAPAAKKLD AEMPNTLFGSVALMYIKQVR RVARKAVLQRELAAKLAKAQ QDLADKQAALVEAERVKAER EAEEARLAAEAEAAAAAEAE AAARAAAEAEAAAAAEAAAE AAAEAAAAAAEAAAEAGEGE AVAEREAAPAEAEAAPAEGE AAPPAEGEGEAQPAQEGSNS SSSSSDSSSSEESKAAAE ; loop_ _Residue_seq_code _Residue_label 1 MET 2 ALA 3 PRO 4 THR 5 GLN 6 ALA 7 GLY 8 HIS 9 ASP 10 THR 11 ALA 12 TYR 13 LEU 14 LYS 15 GLU 16 THR 17 VAL 18 GLY 19 GLU 20 ALA 21 LEU 22 ALA 23 ARG 24 GLY 25 CYS 26 ALA 27 ALA 28 ALA 29 ILE 30 SER 31 ALA 32 GLN 33 PRO 34 ASN 35 ASP 36 PRO 37 VAL 38 GLU 39 TYR 40 LEU 41 GLY 42 LEU 43 TRP 44 LEU 45 LEU 46 LYS 47 TYR 48 VAL 49 LYS 50 ASN 51 ALA 52 GLU 53 VAL 54 GLU 55 GLY 56 ASN 57 PHE 58 TYR 59 ARG 60 GLU 61 ARG 62 GLN 63 GLN 64 ASP 65 LEU 66 GLN 67 LYS 68 LYS 69 LYS 70 ASP 71 ARG 72 LEU 73 VAL 74 LYS 75 GLU 76 ALA 77 GLN 78 SER 79 GLU 80 GLN 81 ALA 82 ALA 83 LYS 84 SER 85 VAL 86 ALA 87 LEU 88 THR 89 ARG 90 LYS 91 GLU 92 ALA 93 ALA 94 ASP 95 ALA 96 LEU 97 ALA 98 LEU 99 VAL 100 THR 101 ALA 102 GLU 103 PRO 104 ARG 105 GLU 106 LEU 107 LEU 108 GLU 109 ALA 110 ALA 111 VAL 112 LYS 113 LEU 114 VAL 115 LYS 116 GLN 117 HIS 118 THR 119 ALA 120 ALA 121 GLY 122 ALA 123 ALA 124 TYR 125 ALA 126 ALA 127 VAL 128 VAL 129 ALA 130 GLU 131 PRO 132 GLU 133 GLU 134 PRO 135 ASP 136 TRP 137 VAL 138 ALA 139 PRO 140 GLU 141 ASP 142 ASP 143 GLU 144 ALA 145 ALA 146 ALA 147 VAL 148 GLU 149 THR 150 GLU 151 ASP 152 GLU 153 ALA 154 ALA 155 GLY 156 GLY 157 ALA 158 ALA 159 LEU 160 ALA 161 GLU 162 GLY 163 GLU 164 GLU 165 PRO 166 PRO 167 PRO 168 GLU 169 PRO 170 GLU 171 PRO 172 GLU 173 PRO 174 GLU 175 ALA 176 ALA 177 PRO 178 GLU 179 ASP 180 GLY 181 GLU 182 GLY 183 ASP 184 ALA 185 PRO 186 ALA 187 PRO 188 LYS 189 ILE 190 PRO 191 ARG 192 PRO 193 VAL 194 ASP 195 TYR 196 SER 197 LYS 198 LYS 199 TYR 200 PHE 201 ALA 202 TYR 203 VAL 204 ALA 205 ALA 206 SER 207 ALA 208 GLY 209 GLN 210 GLU 211 HIS 212 VAL 213 LEU 214 GLU 215 ALA 216 ASP 217 LEU 218 TYR 219 ARG 220 PRO 221 ALA 222 PRO 223 PRO 224 PRO 225 GLU 226 ASP 227 ALA 228 ASP 229 GLU 230 ASP 231 PHE 232 LYS 233 PRO 234 GLU 235 PRO 236 LEU 237 PRO 238 TYR 239 SER 240 PHE 241 ARG 242 VAL 243 LEU 244 ASP 245 GLU 246 LYS 247 LEU 248 PRO 249 MET 250 LEU 251 TYR 252 VAL 253 PRO 254 ASN 255 VAL 256 ALA 257 ALA 258 GLU 259 GLU 260 ARG 261 VAL 262 LYS 263 PHE 264 PHE 265 ARG 266 LYS 267 PHE 268 PRO 269 LYS 270 ILE 271 GLY 272 SER 273 TYR 274 GLN 275 ALA 276 CYS 277 GLY 278 VAL 279 ALA 280 LEU 281 PRO 282 ALA 283 SER 284 GLY 285 GLU 286 PHE 287 LYS 288 ALA 289 LEU 290 LEU 291 ALA 292 ALA 293 ASP 294 THR 295 LEU 296 PHE 297 PRO 298 GLU 299 GLY 300 SER 301 GLY 302 GLN 303 PRO 304 LEU 305 SER 306 ALA 307 ASP 308 ASP 309 ARG 310 ASP 311 PHE 312 VAL 313 TRP 314 GLU 315 VAL 316 SER 317 GLN 318 SER 319 LEU 320 SER 321 ARG 322 ALA 323 LEU 324 GLU 325 ALA 326 VAL 327 GLN 328 ALA 329 ARG 330 ALA 331 ALA 332 GLU 333 ALA 334 LEU 335 ALA 336 ALA 337 THR 338 SER 339 ALA 340 ALA 341 GLU 342 ALA 343 VAL 344 GLU 345 GLU 346 LEU 347 LYS 348 ALA 349 LYS 350 VAL 351 ALA 352 GLU 353 LEU 354 ARG 355 GLU 356 GLN 357 ALA 358 ALA 359 ALA 360 GLU 361 ALA 362 ALA 363 ALA 364 ALA 365 ALA 366 PRO 367 PRO 368 PRO 369 PRO 370 ALA 371 GLU 372 GLY 373 GLU 374 GLU 375 GLY 376 GLU 377 GLY 378 GLU 379 ALA 380 PRO 381 PRO 382 ALA 383 GLU 384 GLU 385 GLU 386 PRO 387 PRO 388 ALA 389 GLU 390 GLU 391 GLU 392 ALA 393 GLU 394 GLU 395 GLU 396 GLU 397 GLU 398 GLU 399 ALA 400 GLU 401 GLU 402 GLY 403 ALA 404 GLU 405 GLU 406 GLY 407 ALA 408 GLU 409 GLU 410 GLY 411 GLU 412 GLU 413 GLY 414 GLU 415 GLU 416 ALA 417 PRO 418 PRO 419 LYS 420 PRO 421 LYS 422 LYS 423 LYS 424 LYS 425 LYS 426 VAL 427 PHE 428 ASN 429 PRO 430 ILE 431 PRO 432 GLY 433 LEU 434 GLN 435 ALA 436 ALA 437 ILE 438 GLU 439 LYS 440 LEU 441 THR 442 ALA 443 ALA 444 ALA 445 GLU 446 ALA 447 ALA 448 THR 449 GLU 450 ALA 451 ASP 452 ALA 453 ARG 454 ALA 455 GLN 456 ALA 457 ALA 458 VAL 459 ALA 460 LEU 461 GLU 462 LYS 463 GLN 464 ALA 465 LEU 466 ASP 467 GLU 468 VAL 469 VAL 470 ALA 471 LEU 472 ALA 473 SER 474 SER 475 HIS 476 SER 477 ASP 478 ALA 479 THR 480 LEU 481 SER 482 SER 483 LEU 484 ARG 485 ASN 486 MET 487 LEU 488 SER 489 VAL 490 PRO 491 GLN 492 GLY 493 THR 494 TYR 495 HIS 496 VAL 497 VAL 498 LYS 499 ALA 500 LEU 501 LEU 502 HIS 503 LEU 504 LEU 505 GLY 506 ARG 507 PRO 508 ALA 509 ALA 510 SER 511 PHE 512 SER 513 THR 514 TRP 515 LYS 516 ARG 517 ALA 518 HIS 519 SER 520 HIS 521 PHE 522 SER 523 PRO 524 ARG 525 LEU 526 PHE 527 GLU 528 ASP 529 MET 530 ALA 531 ALA 532 TYR 533 ASP 534 ALA 535 THR 536 ALA 537 GLU 538 ARG 539 ASP 540 MET 541 ALA 542 VAL 543 TRP 544 GLY 545 ARG 546 VAL 547 ARG 548 SER 549 CYS 550 TYR 551 LYS 552 ALA 553 ALA 554 PRO 555 ALA 556 ALA 557 LYS 558 LYS 559 LEU 560 ASP 561 ALA 562 GLU 563 MET 564 PRO 565 ASN 566 THR 567 LEU 568 PHE 569 GLY 570 SER 571 VAL 572 ALA 573 LEU 574 MET 575 TYR 576 ILE 577 LYS 578 GLN 579 VAL 580 ARG 581 ARG 582 VAL 583 ALA 584 ARG 585 LYS 586 ALA 587 VAL 588 LEU 589 GLN 590 ARG 591 GLU 592 LEU 593 ALA 594 ALA 595 LYS 596 LEU 597 ALA 598 LYS 599 ALA 600 GLN 601 GLN 602 ASP 603 LEU 604 ALA 605 ASP 606 LYS 607 GLN 608 ALA 609 ALA 610 LEU 611 VAL 612 GLU 613 ALA 614 GLU 615 ARG 616 VAL 617 LYS 618 ALA 619 GLU 620 ARG 621 GLU 622 ALA 623 GLU 624 GLU 625 ALA 626 ARG 627 LEU 628 ALA 629 ALA 630 GLU 631 ALA 632 GLU 633 ALA 634 ALA 635 ALA 636 ALA 637 ALA 638 GLU 639 ALA 640 GLU 641 ALA 642 ALA 643 ALA 644 ARG 645 ALA 646 ALA 647 ALA 648 GLU 649 ALA 650 GLU 651 ALA 652 ALA 653 ALA 654 ALA 655 ALA 656 GLU 657 ALA 658 ALA 659 ALA 660 GLU 661 ALA 662 ALA 663 ALA 664 GLU 665 ALA 666 ALA 667 ALA 668 ALA 669 ALA 670 ALA 671 GLU 672 ALA 673 ALA 674 ALA 675 GLU 676 ALA 677 GLY 678 GLU 679 GLY 680 GLU 681 ALA 682 VAL 683 ALA 684 GLU 685 ARG 686 GLU 687 ALA 688 ALA 689 PRO 690 ALA 691 GLU 692 ALA 693 GLU 694 ALA 695 ALA 696 PRO 697 ALA 698 GLU 699 GLY 700 GLU 701 ALA 702 ALA 703 PRO 704 PRO 705 ALA 706 GLU 707 GLY 708 GLU 709 GLY 710 GLU 711 ALA 712 GLN 713 PRO 714 ALA 715 GLN 716 GLU 717 GLY 718 SER 719 ASN 720 SER 721 SER 722 SER 723 SER 724 SER 725 SER 726 ASP 727 SER 728 SER 729 SER 730 SER 731 GLU 732 GLU 733 SER 734 LYS 735 ALA 736 ALA 737 ALA 738 GLU stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-07-08 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value GB AAQ92371 "radial spoke protein 2 [Chlamydomonas reinhardtii]" 100.00 738 100.00 100.00 0.00e+00 GB EDP06497 "radial spoke protein 2 [Chlamydomonas reinhardtii]" 100.00 738 100.00 100.00 0.00e+00 REF XP_001702718 "radial spoke protein 2 [Chlamydomonas reinhardtii]" 100.00 738 100.00 100.00 0.00e+00 SP Q6UBQ3 "RecName: Full=Flagellar radial spoke protein 2" 100.00 738 100.00 100.00 0.00e+00 stop_ save_ ############# # Ligands # ############# save_ATP _Saveframe_category ligand _Mol_type non-polymer _Name_common "ATP (ADENOSINE-5'-TRIPHOSPHATE)" _BMRB_code . _PDB_code ATP _Molecular_mass 507.181 _Mol_charge 0 _Mol_paramagnetic no _Mol_aromatic yes _Details ; Information obtained from PDB's Chemical Component Dictionary at http://wwpdb-remediation.rutgers.edu/downloads.html Downloaded on Tue Jun 9 13:00:26 2009 ; loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons C1' C1' C . 0 . ? C2 C2 C . 0 . ? C2' C2' C . 0 . ? C3' C3' C . 0 . ? C4 C4 C . 0 . ? C4' C4' C . 0 . ? C5 C5 C . 0 . ? C5' C5' C . 0 . ? C6 C6 C . 0 . ? C8 C8 C . 0 . ? H1' H1' H . 0 . ? H2 H2 H . 0 . ? H2' H2' H . 0 . ? H3' H3' H . 0 . ? H4' H4' H . 0 . ? H5'1 H5'1 H . 0 . ? H5'2 H5'2 H . 0 . ? H8 H8 H . 0 . ? HN61 HN61 H . 0 . ? HN62 HN62 H . 0 . ? HO2' HO2' H . 0 . ? HO3' HO3' H . 0 . ? HOA2 HOA2 H . 0 . ? HOB2 HOB2 H . 0 . ? HOG2 HOG2 H . 0 . ? HOG3 HOG3 H . 0 . ? N1 N1 N . 0 . ? N3 N3 N . 0 . ? N6 N6 N . 0 . ? N7 N7 N . 0 . ? N9 N9 N . 0 . ? O1A O1A O . 0 . ? O1B O1B O . 0 . ? O1G O1G O . 0 . ? O2' O2' O . 0 . ? O2A O2A O . 0 . ? O2B O2B O . 0 . ? O2G O2G O . 0 . ? O3' O3' O . 0 . ? O3A O3A O . 0 . ? O3B O3B O . 0 . ? O3G O3G O . 0 . ? O4' O4' O . 0 . ? O5' O5' O . 0 . ? PA PA P . 0 . ? PB PB P . 0 . ? PG PG P . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name DOUB PG O1G ? ? SING PG O2G ? ? SING PG O3G ? ? SING PG O3B ? ? SING O2G HOG2 ? ? SING O3G HOG3 ? ? DOUB PB O1B ? ? SING PB O2B ? ? SING PB O3B ? ? SING PB O3A ? ? SING O2B HOB2 ? ? DOUB PA O1A ? ? SING PA O2A ? ? SING PA O3A ? ? SING PA O5' ? ? SING O2A HOA2 ? ? SING O5' C5' ? ? SING C5' C4' ? ? SING C5' H5'1 ? ? SING C5' H5'2 ? ? SING C4' O4' ? ? SING C4' C3' ? ? SING C4' H4' ? ? SING O4' C1' ? ? SING C3' O3' ? ? SING C3' C2' ? ? SING C3' H3' ? ? SING O3' HO3' ? ? SING C2' O2' ? ? SING C2' C1' ? ? SING C2' H2' ? ? SING O2' HO2' ? ? SING C1' N9 ? ? SING C1' H1' ? ? SING N9 C8 ? ? SING N9 C4 ? ? DOUB C8 N7 ? ? SING C8 H8 ? ? SING N7 C5 ? ? SING C5 C6 ? ? DOUB C5 C4 ? ? SING C6 N6 ? ? DOUB C6 N1 ? ? SING N6 HN61 ? ? SING N6 HN62 ? ? SING N1 C2 ? ? DOUB C2 N3 ? ? SING C2 H2 ? ? SING N3 C4 ? ? stop_ _Mol_thiol_state 'not present' _Sequence_homology_query_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $RSP2 'Green algae' 3055 Eukaryota Viridiplantae Chlamydomonas reinhardtii stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Variant _Vector_name $RSP2 'recombinant technology' . Escherichia coli BL21 DE3 pET stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium chloride' 200 mM 'natural abundance' 'sodium phosphate' 20 mM 'natural abundance' $RSP2 10 uM 'natural abundance' $ATP 1 mM 'natural abundance' GTP 1 mM 'natural abundance' CMP 1 mM 'natural abundance' PCG 1 mM 'natural abundance' cCMP 1 mM 'natural abundance' AMP 1 mM 'natural abundance' D2O 100 % 'natural abundance' stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling 'sodium chloride' 200 mM 'natural abundance' 'sodium phosphate' 20 mM 'natural abundance' $RSP2 10 uM 'natural abundance' $ATP 1 mM 'natural abundance' GTP 1 mM 'natural abundance' CMP 1 mM 'natural abundance' PCG 1 mM 'natural abundance' cCMP 1 mM 'natural abundance' AMP 1 mM 'natural abundance' MG 4 mM 'natural abundance' D2O 100 % 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_MestRe-C _Saveframe_category software _Name MestRe-C _Version . loop_ _Vendor _Address _Electronic_address '(University of Santiago, Spain)' . . stop_ loop_ _Task processing stop_ _Details . save_ save_SigmaPlot _Saveframe_category software _Name SigmaPlot _Version 8.0 loop_ _Vendor _Address _Electronic_address 'aspire software international' . . stop_ loop_ _Task 'data analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model 300 _Field_strength 121.5 _Details 'Madison NMR facility' save_ ############################# # NMR applied experiments # ############################# save_1D_STD_1H_NMR_1 _Saveframe_category NMR_applied_experiment _Experiment_name '1D STD 1H NMR' _Sample_label $sample_1 save_ save_1D_STD_1H_NMR_2 _Saveframe_category NMR_applied_experiment _Experiment_name '1D STD 1H NMR' _Sample_label $sample_2 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 7.4 . pH pressure 1 . atm temperature 277 . K stop_ save_