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New page: left|200px<br /><applet load="1djn" size="450" color="white" frame="true" align="right" spinBox="true" caption="1djn, resolution 2.2Å" /> '''STRUCTURAL AND BIOCHE...
 
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[[Image:1djn.gif|left|200px]]<br /><applet load="1djn" size="450" color="white" frame="true" align="right" spinBox="true"
caption="1djn, resolution 2.2&Aring;" />
'''STRUCTURAL AND BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT WILD TYPE TRIMETHYLAMINE DEHYDROGENASE FROM METHYLOPHILUS METHYLOTROPHUS (SP. W3A1)'''<br />


==Overview==
==STRUCTURAL AND BIOCHEMICAL CHARACTERIZATION OF RECOMBINANT WILD TYPE TRIMETHYLAMINE DEHYDROGENASE FROM METHYLOPHILUS METHYLOTROPHUS (SP. W3A1)==
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.
<StructureSection load='1djn' size='340' side='right'caption='[[1djn]], [[Resolution|resolution]] 2.20&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[1djn]] 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=1DJN OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1DJN FirstGlance]. <br>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.2&#8491;</td></tr>
<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></td></tr>
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=1djn FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1djn OCA], [https://pdbe.org/1djn PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1djn RCSB], [https://www.ebi.ac.uk/pdbsum/1djn PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1djn ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/DHTM_METME DHTM_METME]
== Evolutionary Conservation ==
[[Image:Consurf_key_small.gif|200px|right]]
Check<jmol>
  <jmolCheckbox>
    <scriptWhenChecked>; select protein; define ~consurf_to_do selected; consurf_initial_scene = true; script "/wiki/ConSurf/dj/1djn_consurf.spt"</scriptWhenChecked>
    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview03.spt</scriptWhenUnchecked>
    <text>to colour the structure by Evolutionary Conservation</text>
  </jmolCheckbox>
</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=1djn ConSurf].
<div style="clear:both"></div>
<div style="background-color:#fffaf0;">
== Publication Abstract from PubMed ==
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.


==About this Structure==
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:10869173<ref>PMID:10869173</ref>
1DJN is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Methylophilus_methylotrophus_w3a1 Methylophilus methylotrophus w3a1] with SF4, FMN and ADP as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [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] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1DJN OCA].


==Reference==
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
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://ispc.weizmann.ac.il//pmbin/getpm?pmid=10869173 10869173]
</div>
[[Category: Methylophilus methylotrophus w3a1]]
<div class="pdbe-citations 1djn" style="background-color:#fffaf0;"></div>
[[Category: Single protein]]
== References ==
[[Category: Trimethylamine dehydrogenase]]
<references/>
[[Category: Barton, J.D.]]
__TOC__
[[Category: Basran, J.]]
</StructureSection>
[[Category: Chen, Z.W.]]
[[Category: Large Structures]]
[[Category: Lian, L.Y.]]
[[Category: Methylophilus methylotrophus W3A1]]
[[Category: Mathews, F.S.]]
[[Category: Barton JD]]
[[Category: Scrutton, N.S.]]
[[Category: Basran J]]
[[Category: Sutcliffe, M.J.]]
[[Category: Chen Z-W]]
[[Category: Trickey, P.]]
[[Category: Lian L-Y]]
[[Category: ADP]]
[[Category: Mathews FS]]
[[Category: FMN]]
[[Category: Scrutton NS]]
[[Category: SF4]]
[[Category: Sutcliffe MJ]]
[[Category: electron transfer]]
[[Category: Trickey P]]
[[Category: iron-sulfur flavoprotein]]
[[Category: oxidoreductase]]
 
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