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[[Image:1a5b.gif|left|200px]]<br />
<applet load="1a5b" size="450" color="white" frame="true" align="right" spinBox="true"
caption="1a5b, resolution 2.0&Aring;" />
'''CRYO-CRYSTALLOGRAPHY OF A TRUE SUBSTRATE, INDOLE-3-GLYCEROL PHOSPHATE, BOUND TO A MUTANT (ALPHA D60N) TRYPTOPHAN SYNTHASE ALPHA2BETA2 COMPLEX REVEALS THE CORRECT ORIENTATION OF ACTIVE SITE ALPHA GLU 49'''<br />


==Overview==
==CRYO-CRYSTALLOGRAPHY OF A TRUE SUBSTRATE, INDOLE-3-GLYCEROL PHOSPHATE, BOUND TO A MUTANT (ALPHA D60N) TRYPTOPHAN SYNTHASE ALPHA2BETA2 COMPLEX REVEALS THE CORRECT ORIENTATION OF ACTIVE SITE ALPHA GLU 49==
The reversible cleavage of indole-3-glycerol by the alpha-subunit of, tryptophan synthase has been proposed to be catalyzed by alphaGlu49 and, alphaAsp60. Although previous x-ray crystallographic structures of the, tryptophan synthase alpha2beta2 complex showed an interaction between the, carboxylate of alphaAsp60 and the bound inhibitor indole-3-propanol, phosphate, the carboxylate of alphaGlu49 was too distant to play its, proposed role. To clarify the structural and functional roles of, alphaGlu49, we have determined crystal structures of a mutant (alphaD60N), alpha2beta2 complex in the presence and absence of the true substrate, indole-3-glycerol phosphate. The enzyme in the crystal cleaves, indole-3-glycerol phosphate very slowly at room temperature but not under, cryo-conditions of 95 K. The structure of the complex with the true, substrate obtained by cryo-crystallography reveals that indole-3-glycerol, phosphate and indole-3-propanol phosphate have similar binding modes but, different torsion angles. Most importantly, the side chain of alphaGlu49, interacts with 3-hydroxyl group of indole-3-glycerol phosphate as, proposed. The movement of the side chain of alphaGlu49 into an extended, conformation upon binding the true substrate provides evidence for an, induced fit mechanism. Our results demonstrate how cryo-crystallography, and mutagenesis can provide insight into enzyme mechanism.
<StructureSection load='1a5b' size='340' side='right'caption='[[1a5b]], [[Resolution|resolution]] 2.00&Aring;' scene=''>
== Structural highlights ==
<table><tr><td colspan='2'>[[1a5b]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Salmonella_enterica_subsp._enterica_serovar_Typhimurium Salmonella enterica subsp. enterica serovar Typhimurium]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1A5B OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=1A5B 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&#8491;</td></tr>
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=IGP:INDOLE-3-GLYCEROL+PHOSPHATE'>IGP</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=PLP:PYRIDOXAL-5-PHOSPHATE'>PLP</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=1a5b FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1a5b OCA], [https://pdbe.org/1a5b PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=1a5b RCSB], [https://www.ebi.ac.uk/pdbsum/1a5b PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=1a5b ProSAT]</span></td></tr>
</table>
== Function ==
[https://www.uniprot.org/uniprot/TRPA_SALTY TRPA_SALTY] The alpha subunit is responsible for the aldol cleavage of indoleglycerol phosphate to indole and glyceraldehyde 3-phosphate.
== 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/a5/1a5b_consurf.spt"</scriptWhenChecked>
    <scriptWhenUnchecked>script /wiki/extensions/Proteopedia/spt/initialview01.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=1a5b ConSurf].
<div style="clear:both"></div>


==About this Structure==
==See Also==
1A5B is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Salmonella_typhimurium Salmonella typhimurium] with K, IGP and PLP as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Tryptophan_synthase Tryptophan synthase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=4.2.1.20 4.2.1.20] Structure known Active Sites: S1 and S2. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1A5B OCA].
*[[Tryptophan synthase 3D structures|Tryptophan synthase 3D structures]]
 
__TOC__
==Reference==
</StructureSection>
Cryo-crystallography of a true substrate, indole-3-glycerol phosphate, bound to a mutant (alphaD60N) tryptophan synthase alpha2beta2 complex reveals the correct orientation of active site alphaGlu49., Rhee S, Miles EW, Davies DR, J Biol Chem. 1998 Apr 10;273(15):8553-5. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=9535826 9535826]
[[Category: Large Structures]]
[[Category: Protein complex]]
[[Category: Salmonella enterica subsp. enterica serovar Typhimurium]]
[[Category: Salmonella typhimurium]]
[[Category: Davies DR]]
[[Category: Tryptophan synthase]]
[[Category: Miles EW]]
[[Category: Davies, D.R.]]
[[Category: Rhee S]]
[[Category: Miles, E.W.]]
[[Category: Rhee, S.]]
[[Category: IGP]]
[[Category: K]]
[[Category: PLP]]
[[Category: carbon-oxygen lyase]]
[[Category: mutation at position 60 (asp --> asn) in the a-subunit]]
[[Category: true substrate indole-3-glycerol phosphate in the a-subunit]]
 
''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov  5 15:44:51 2007''

Latest revision as of 18:21, 13 March 2024

CRYO-CRYSTALLOGRAPHY OF A TRUE SUBSTRATE, INDOLE-3-GLYCEROL PHOSPHATE, BOUND TO A MUTANT (ALPHA D60N) TRYPTOPHAN SYNTHASE ALPHA2BETA2 COMPLEX REVEALS THE CORRECT ORIENTATION OF ACTIVE SITE ALPHA GLU 49CRYO-CRYSTALLOGRAPHY OF A TRUE SUBSTRATE, INDOLE-3-GLYCEROL PHOSPHATE, BOUND TO A MUTANT (ALPHA D60N) TRYPTOPHAN SYNTHASE ALPHA2BETA2 COMPLEX REVEALS THE CORRECT ORIENTATION OF ACTIVE SITE ALPHA GLU 49

Structural highlights

1a5b is a 2 chain structure with sequence from Salmonella enterica subsp. enterica serovar Typhimurium. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:X-ray diffraction, Resolution 2Å
Ligands:, ,
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

TRPA_SALTY The alpha subunit is responsible for the aldol cleavage of indoleglycerol phosphate to indole and glyceraldehyde 3-phosphate.

Evolutionary Conservation

Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.

See Also

1a5b, resolution 2.00Å

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