1tsv
From Proteopedia
(New page: 200px<br /><applet load="1tsv" size="450" color="white" frame="true" align="right" spinBox="true" caption="1tsv, resolution 2.9Å" /> '''THYMIDYLATE SYNTHASE ...) |
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| - | [[Image:1tsv.jpg|left|200px]]<br /><applet load="1tsv" size=" | + | [[Image:1tsv.jpg|left|200px]]<br /><applet load="1tsv" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1tsv, resolution 2.9Å" /> | caption="1tsv, resolution 2.9Å" /> | ||
'''THYMIDYLATE SYNTHASE R179A MUTANT'''<br /> | '''THYMIDYLATE SYNTHASE R179A MUTANT'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Invariant arginine 179, one of four arginines that are conserved in all | + | Invariant arginine 179, one of four arginines that are conserved in all thymidylate synthases (TS) and that bind the phosphate moiety of the substrate 2'-deoxyuridine-5'-monophosphate (dUMP), can be altered even to a negatively charged glutamic acid with little effect on kcat. In the mutant structures, ordered water or the other phosphate-binding arginines compensate for the hydrogen bonds made by Arg179 in the wild-type enzyme and there is almost no change in the conformation or binding site of dUMP. Correlation of dUMP Kds for TS R179A and TS R179K with the structures of their binary complexes shows, that the positive charge on Arg179 contributes significantly to dUMP binding affinity. kcat/K(m) for dUMP measures the rate of dUMP binding to TS during the ordered bi-substrate reaction, and in the ternary complex dUMP provides a binding surface for the cofactor. kcat/K(m) reflects the ability of the enzyme to accept a properly oriented dUMP for catalysis and is less sensitive than is Kd to the changes in electrostatics at the phosphate binding site. |
==About this Structure== | ==About this Structure== | ||
| - | 1TSV is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Lactobacillus_casei Lactobacillus casei] with UMP as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Thymidylate_synthase Thymidylate synthase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.1.1.45 2.1.1.45] Full crystallographic information is available from [http:// | + | 1TSV is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Lactobacillus_casei Lactobacillus casei] with <scene name='pdbligand=UMP:'>UMP</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Thymidylate_synthase Thymidylate synthase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.1.1.45 2.1.1.45] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1TSV OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Thymidylate synthase]] | [[Category: Thymidylate synthase]] | ||
[[Category: Finer-Moore, J.]] | [[Category: Finer-Moore, J.]] | ||
| - | [[Category: Stroud, R | + | [[Category: Stroud, R M.]] |
[[Category: UMP]] | [[Category: UMP]] | ||
[[Category: methyltransferase]] | [[Category: methyltransferase]] | ||
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[[Category: transferase]] | [[Category: transferase]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:17:00 2008'' |
Revision as of 13:17, 21 February 2008
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THYMIDYLATE SYNTHASE R179A MUTANT
Overview
Invariant arginine 179, one of four arginines that are conserved in all thymidylate synthases (TS) and that bind the phosphate moiety of the substrate 2'-deoxyuridine-5'-monophosphate (dUMP), can be altered even to a negatively charged glutamic acid with little effect on kcat. In the mutant structures, ordered water or the other phosphate-binding arginines compensate for the hydrogen bonds made by Arg179 in the wild-type enzyme and there is almost no change in the conformation or binding site of dUMP. Correlation of dUMP Kds for TS R179A and TS R179K with the structures of their binary complexes shows, that the positive charge on Arg179 contributes significantly to dUMP binding affinity. kcat/K(m) for dUMP measures the rate of dUMP binding to TS during the ordered bi-substrate reaction, and in the ternary complex dUMP provides a binding surface for the cofactor. kcat/K(m) reflects the ability of the enzyme to accept a properly oriented dUMP for catalysis and is less sensitive than is Kd to the changes in electrostatics at the phosphate binding site.
About this Structure
1TSV is a Single protein structure of sequence from Lactobacillus casei with as ligand. Active as Thymidylate synthase, with EC number 2.1.1.45 Full crystallographic information is available from OCA.
Reference
Contribution of a salt bridge to binding affinity and dUMP orientation to catalytic rate: mutation of a substrate-binding arginine in thymidylate synthase., Finer-Moore JS, Fauman EB, Morse RJ, Santi DV, Stroud RM, Protein Eng. 1996 Jan;9(1):69-75. PMID:9053905
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