1xql

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[[Image:1xql.gif|left|200px]]<br /><applet load="1xql" size="350" color="white" frame="true" align="right" spinBox="true"
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[[Image:1xql.gif|left|200px]]
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caption="1xql, resolution 1.80&Aring;" />
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'''Effect of a Y265F Mutant on the Transamination Based Cycloserine Inactivation of Alanine Racemase'''<br />
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{{Structure
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|PDB= 1xql |SIZE=350|CAPTION= <scene name='initialview01'>1xql</scene>, resolution 1.80&Aring;
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|SITE=
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|LIGAND= <scene name='pdbligand=PMP:4'-DEOXY-4'-AMINOPYRIDOXAL-5'-PHOSPHATE'>PMP</scene>, <scene name='pdbligand=PMH:(5-HYDROXY-4-{[(3-HYDROXYISOXAZOL-4-YL)AMINO]METHYL}-6-METHYLPYRIDIN-3-YL)METHYL+DIHYDROGEN+PHOSPHATE'>PMH</scene>, <scene name='pdbligand=PLP:PYRIDOXAL-5'-PHOSPHATE'>PLP</scene>, <scene name='pdbligand=4AX:(R)-4-AMINO-ISOXAZOLIDIN-3-ONE'>4AX</scene> and <scene name='pdbligand=ACY:ACETIC ACID'>ACY</scene>
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|ACTIVITY= [http://en.wikipedia.org/wiki/Alanine_racemase Alanine racemase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.1.1.1 5.1.1.1]
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|GENE= alr, dal ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1422 Geobacillus stearothermophilus])
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}}
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'''Effect of a Y265F Mutant on the Transamination Based Cycloserine Inactivation of Alanine Racemase'''
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==Overview==
==Overview==
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==About this Structure==
==About this Structure==
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1XQL is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Geobacillus_stearothermophilus Geobacillus stearothermophilus] with <scene name='pdbligand=PMP:'>PMP</scene>, <scene name='pdbligand=PMH:'>PMH</scene>, <scene name='pdbligand=PLP:'>PLP</scene>, <scene name='pdbligand=4AX:'>4AX</scene> and <scene name='pdbligand=ACY:'>ACY</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Alanine_racemase Alanine racemase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=5.1.1.1 5.1.1.1] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1XQL OCA].
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1XQL is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Geobacillus_stearothermophilus Geobacillus stearothermophilus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1XQL OCA].
==Reference==
==Reference==
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Effect of a Y265F mutant on the transamination-based cycloserine inactivation of alanine racemase., Fenn TD, Holyoak T, Stamper GF, Ringe D, Biochemistry. 2005 Apr 12;44(14):5317-27. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=15807525 15807525]
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Effect of a Y265F mutant on the transamination-based cycloserine inactivation of alanine racemase., Fenn TD, Holyoak T, Stamper GF, Ringe D, Biochemistry. 2005 Apr 12;44(14):5317-27. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15807525 15807525]
[[Category: Alanine racemase]]
[[Category: Alanine racemase]]
[[Category: Geobacillus stearothermophilus]]
[[Category: Geobacillus stearothermophilus]]
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[[Category: tim barrel]]
[[Category: tim barrel]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:57:35 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 15:13:15 2008''

Revision as of 13:13, 20 March 2008

Image:1xql.gif


PDB ID 1xql

Drag the structure with the mouse to rotate
, resolution 1.80Å
Ligands: , , , and
Gene: alr, dal (Geobacillus stearothermophilus)
Activity: Alanine racemase, with EC number 5.1.1.1
Coordinates: save as pdb, mmCIF, xml



Effect of a Y265F Mutant on the Transamination Based Cycloserine Inactivation of Alanine Racemase


Overview

The requirement for d-alanine in the peptidoglycan layer of bacterial cell walls is fulfilled in part by alanine racemase (EC 5.1.1.1), a pyridoxal 5'-phosphate (PLP)-assisted enzyme. The enzyme utilizes two antiparallel bases focused at the C(alpha) position and oriented perpendicular to the PLP ring to facilitate the equilibration of alanine enantiomers. Understanding how this two-base system is utilized and controlled to yield reaction specificity is therefore a potential means for designing antibiotics. Cycloserine is a known alanine racemase suicide substrate, although its mechanism of inactivation is based on transaminase chemistry. Here we characterize the effects of a Y265F mutant (Tyr265 acts as the catalytic base in the l-isomer case) of Bacillus stearothermophilus alanine racemase on cycloserine inactivation. The Y265F mutant reduces racemization activity 1600-fold [Watanabe, A., Yoshimura, T., Mikami, B., and Esaki, N. (1999) J. Biochem. 126, 781-786] and only leads to formation of the isoxazole end product (the result of the transaminase pathway) in the case of d-cycloserine, in contrast to results obtained using the wild-type enzyme. l-Cycloserine, on the other hand, utilizes a number of alternative pathways in the absence of Y265, emphasizing the importance of Y265 in both the inactivation and racemization pathway. In combination with the kinetics of inactivation, these results suggest roles for each of the two catalytic bases in racemization and inactivation, as well as the importance of Y265 in "steering" the chemistry to favor one pathway over another.

About this Structure

1XQL is a Single protein structure of sequence from Geobacillus stearothermophilus. Full crystallographic information is available from OCA.

Reference

Effect of a Y265F mutant on the transamination-based cycloserine inactivation of alanine racemase., Fenn TD, Holyoak T, Stamper GF, Ringe D, Biochemistry. 2005 Apr 12;44(14):5317-27. PMID:15807525

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