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| - | [[Image:1djq.gif|left|200px]] | |
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| - | {{Structure
| + | ==STRUCTURAL AND BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT C30A MUTANT OF TRIMETHYLAMINE DEHYDROGENASE FROM METHYLOPHILUS METHYLOTROPHUS (SP. W3A1)== |
| - | |PDB= 1djq |SIZE=350|CAPTION= <scene name='initialview01'>1djq</scene>, resolution 2.2Å
| + | <StructureSection load='1djq' size='340' side='right'caption='[[1djq]], [[Resolution|resolution]] 2.20Å' scene=''> |
| - | |SITE=
| + | == Structural highlights == |
| - | |LIGAND= <scene name='pdbligand=ADP:ADENOSINE-5'-DIPHOSPHATE'>ADP</scene>, <scene name='pdbligand=FMN:FLAVIN+MONONUCLEOTIDE'>FMN</scene>, <scene name='pdbligand=SF4:IRON/SULFUR+CLUSTER'>SF4</scene> | + | <table><tr><td colspan='2'>[[1djq]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Methylophilus_methylotrophus_W3A1 Methylophilus methylotrophus W3A1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1DJQ OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1DJQ FirstGlance]. <br> |
| - | |ACTIVITY= <span class='plainlinks'>[http://en.wikipedia.org/wiki/Trimethylamine_dehydrogenase Trimethylamine dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.5.8.2 1.5.8.2] </span>
| + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.2Å</td></tr> |
| - | |GENE=
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ADP:ADENOSINE-5-DIPHOSPHATE'>ADP</scene>, <scene name='pdbligand=FMN:FLAVIN+MONONUCLEOTIDE'>FMN</scene>, <scene name='pdbligand=SF4:IRON/SULFUR+CLUSTER'>SF4</scene></td></tr> |
| - | |DOMAIN=
| + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1djq FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1djq OCA], [https://pdbe.org/1djq PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1djq RCSB], [https://www.ebi.ac.uk/pdbsum/1djq PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1djq ProSAT]</span></td></tr> |
| - | |RELATEDENTRY=[[2tmd|2TMD]], [[1djn|1DJN]]
| + | </table> |
| - | |RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1djq FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1djq OCA], [http://www.ebi.ac.uk/pdbsum/1djq PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1djq RCSB]</span>
| + | == Function == |
| - | }}
| + | [https://www.uniprot.org/uniprot/DHTM_METME DHTM_METME] |
| - | | + | == Evolutionary Conservation == |
| - | '''STRUCTURAL AND BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT C30A MUTANT OF TRIMETHYLAMINE DEHYDROGENASE FROM METHYLOPHILUS METHYLOTROPHUS (SP. W3A1)'''
| + | [[Image:Consurf_key_small.gif|200px|right]] |
| - | | + | Check<jmol> |
| - | | + | <jmolCheckbox> |
| - | ==Overview== | + | <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/dj/1djq_consurf.spt"</scriptWhenChecked> |
| - | Trimethylamine dehydrogenase (TMADH) is an iron-sulfur flavoprotein that catalyzes the oxidative demethylation of trimethylamine to form dimethylamine and formaldehyde. It contains a unique flavin, in the form of a 6-S-cysteinyl FMN, which is bent by approximately 25 degrees along the N5-N10 axis of the flavin isoalloxazine ring. This unusual conformation is thought to modulate the properties of the flavin to facilitate catalysis, and has been postulated to be the result of covalent linkage to Cys-30 at the flavin C6 atom. We report here the crystal structures of recombinant wild-type and the C30A mutant TMADH enzymes, both determined at 2.2 A resolution. Combined crystallographic and NMR studies reveal the presence of inorganic phosphate in the FMN binding site in the deflavo fraction of both recombinant wild-type and C30A proteins. The presence of tightly bound inorganic phosphate in the recombinant enzymes explains the inability to reconstitute the deflavo forms of the recombinant wild-type and C30A enzymes that are generated in vivo. The active site structure and flavin conformation in C30A TMADH are identical to those in recombinant and native TMADH, thus revealing that, contrary to expectation, the 6-S-cysteinyl FMN link is not responsible for the 25 degrees butterfly bending along the N5-N10 axis of the flavin in TMADH. Computational quantum chemistry studies strongly support the proposed role of the butterfly bend in modulating the redox properties of the flavin. Solution studies reveal major differences in the kinetic behavior of the wild-type and C30A proteins. Computational studies reveal a hitherto, unrecognized, contribution made by the S(gamma) atom of Cys-30 to substrate binding, and a role for Cys-30 in the optimal geometrical alignment of substrate with the 6-S-cysteinyl FMN in the enzyme active site.
| + | <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.spt</scriptWhenUnchecked> |
| - | | + | <text>to colour the structure by Evolutionary Conservation</text> |
| - | ==About this Structure== | + | </jmolCheckbox> |
| - | 1DJQ is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Methylophilus_methylotrophus_w3a1 Methylophilus methylotrophus w3a1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1DJQ OCA].
| + | </jmol>, as determined by [http://consurfdb.tau.ac.il/ ConSurfDB]. You may read the [[Conservation%2C_Evolutionary|explanation]] of the method and the full data available from [http://bental.tau.ac.il/new_ConSurfDB/main_output.php?pdb_ID=1djq ConSurf]. |
| - | | + | <div style="clear:both"></div> |
| - | ==Reference==
| + | __TOC__ |
| - | Structural and biochemical characterization of recombinant wild type and a C30A mutant of trimethylamine dehydrogenase from methylophilus methylotrophus (sp. W(3)A(1))., Trickey P, Basran J, Lian LY, Chen Z, Barton JD, Sutcliffe MJ, Scrutton NS, Mathews FS, Biochemistry. 2000 Jul 4;39(26):7678-88. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/10869173 10869173]
| + | </StructureSection> |
| - | [[Category: Methylophilus methylotrophus w3a1]]
| + | [[Category: Large Structures]] |
| - | [[Category: Single protein]] | + | [[Category: Methylophilus methylotrophus W3A1]] |
| - | [[Category: Trimethylamine dehydrogenase]] | + | [[Category: Barton JD]] |
| - | [[Category: Barton, J D.]] | + | [[Category: Basran J]] |
| - | [[Category: Basran, J.]] | + | [[Category: Chen Z-W]] |
| - | [[Category: Chen, Z W.]] | + | [[Category: Lian L-Y]] |
| - | [[Category: Lian, L Y.]] | + | [[Category: Mathews FS]] |
| - | [[Category: Mathews, F S.]] | + | [[Category: Scrutton NS]] |
| - | [[Category: Scrutton, N S.]] | + | [[Category: Sutcliffe MJ]] |
| - | [[Category: Sutcliffe, M J.]] | + | [[Category: Trickey P]] |
| - | [[Category: Trickey, P.]] | + | |
| - | [[Category: electron transfer]]
| + | |
| - | [[Category: iron-sulfur flavoprotein]]
| + | |
| - | [[Category: oxidoreductase]]
| + | |
| - | | + | |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 19:42:50 2008''
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