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1a5b

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[[Image:1a5b.gif|left|200px]]
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{{Seed}}
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[[Image:1a5b.png|left|200px]]
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{{STRUCTURE_1a5b| PDB=1a5b | SCENE= }}
{{STRUCTURE_1a5b| PDB=1a5b | SCENE= }}
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'''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'''
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===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===
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==Overview==
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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.
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(as it appears on PubMed at http://www.pubmed.gov), where 9535826 is the PubMed ID number.
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{{ABSTRACT_PUBMED_9535826}}
==About this Structure==
==About this Structure==
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[[Category: Carbon-oxygen lyase]]
[[Category: Carbon-oxygen lyase]]
[[Category: True substrate indole-3-glycerol phosphate in the a-subunit]]
[[Category: True substrate indole-3-glycerol phosphate in the a-subunit]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 09:50:04 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jun 30 16:09:52 2008''

Revision as of 13:09, 30 June 2008

Image:1a5b.png

Template:STRUCTURE 1a5b

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

Template:ABSTRACT PUBMED 9535826

About this Structure

1A5B is a Protein complex structure of sequences from Salmonella typhimurium. Full crystallographic information is available from OCA.

Reference

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:9535826

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