2g37
From Proteopedia
(New page: 200px<br /><applet load="2g37" size="350" color="white" frame="true" align="right" spinBox="true" caption="2g37, resolution 2.000Å" /> '''Structure of Thermu...) |
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==Overview== | ==Overview== | ||
| - | Proline dehydrogenase (PRODH) and pyrroline-5-carboxylate dehydrogenase | + | Proline dehydrogenase (PRODH) and Delta(1)-pyrroline-5-carboxylate dehydrogenase (P5CDH) catalyze the two-step oxidation of proline to glutamate. They are distinct monofunctional enzymes in all eukaryotes and some bacteria but are fused into bifunctional enzymes known as proline utilization A (PutA) in other bacteria. Here we report the first structure and biochemical data for a monofunctional PRODH. The 2.0-A resolution structure of Thermus thermophilus PRODH reveals a distorted (betaalpha)(8) barrel catalytic core domain and a hydrophobic alpha-helical domain located above the carboxyl-terminal ends of the strands of the barrel. Although the catalytic core is similar to that of the PutA PRODH domain, the FAD conformation of T. thermophilus PRODH is remarkably different and likely reflects unique requirements for membrane association and communication with P5CDH. Also, the FAD of T. thermophilus PRODH is highly solvent-exposed compared with PutA due to a 4-A shift of helix 8. Structure-based sequence analysis of the PutA/PRODH family led us to identify nine conserved motifs involved in cofactor and substrate recognition. Biochemical studies show that the midpoint potential of the FAD is -75 mV and the kinetic parameters for proline are K(m) = 27 mm and k(cat) = 13 s(-1). 3,4-Dehydro-l-proline was found to be an efficient substrate, and l-tetrahydro-2-furoic acid is a competitive inhibitor (K(I) = 1.0 mm). Finally, we demonstrate that T. thermophilus PRODH reacts with O(2) producing superoxide. This is significant because superoxide production underlies the role of human PRODH in p53-mediated apoptosis, implying commonalities between eukaryotic and bacterial monofunctional PRODHs. |
==About this Structure== | ==About this Structure== | ||
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==Reference== | ==Reference== | ||
| - | Structure and kinetics of monofunctional proline dehydrogenase from | + | Structure and kinetics of monofunctional proline dehydrogenase from Thermus thermophilus., White TA, Krishnan N, Becker DF, Tanner JJ, J Biol Chem. 2007 May 11;282(19):14316-27. Epub 2007 Mar 7. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17344208 17344208] |
[[Category: Proline dehydrogenase]] | [[Category: Proline dehydrogenase]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Thermus thermophilus]] | [[Category: Thermus thermophilus]] | ||
| - | [[Category: Tanner, J | + | [[Category: Tanner, J J.]] |
| - | [[Category: White, T | + | [[Category: White, T A.]] |
[[Category: FAD]] | [[Category: FAD]] | ||
[[Category: MPD]] | [[Category: MPD]] | ||
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[[Category: flavoenzyme]] | [[Category: flavoenzyme]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:27:40 2008'' |
Revision as of 15:27, 21 February 2008
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Structure of Thermus thermophilus L-proline dehydrogenase
Overview
Proline dehydrogenase (PRODH) and Delta(1)-pyrroline-5-carboxylate dehydrogenase (P5CDH) catalyze the two-step oxidation of proline to glutamate. They are distinct monofunctional enzymes in all eukaryotes and some bacteria but are fused into bifunctional enzymes known as proline utilization A (PutA) in other bacteria. Here we report the first structure and biochemical data for a monofunctional PRODH. The 2.0-A resolution structure of Thermus thermophilus PRODH reveals a distorted (betaalpha)(8) barrel catalytic core domain and a hydrophobic alpha-helical domain located above the carboxyl-terminal ends of the strands of the barrel. Although the catalytic core is similar to that of the PutA PRODH domain, the FAD conformation of T. thermophilus PRODH is remarkably different and likely reflects unique requirements for membrane association and communication with P5CDH. Also, the FAD of T. thermophilus PRODH is highly solvent-exposed compared with PutA due to a 4-A shift of helix 8. Structure-based sequence analysis of the PutA/PRODH family led us to identify nine conserved motifs involved in cofactor and substrate recognition. Biochemical studies show that the midpoint potential of the FAD is -75 mV and the kinetic parameters for proline are K(m) = 27 mm and k(cat) = 13 s(-1). 3,4-Dehydro-l-proline was found to be an efficient substrate, and l-tetrahydro-2-furoic acid is a competitive inhibitor (K(I) = 1.0 mm). Finally, we demonstrate that T. thermophilus PRODH reacts with O(2) producing superoxide. This is significant because superoxide production underlies the role of human PRODH in p53-mediated apoptosis, implying commonalities between eukaryotic and bacterial monofunctional PRODHs.
About this Structure
2G37 is a Single protein structure of sequence from Thermus thermophilus with and as ligands. Active as Proline dehydrogenase, with EC number 1.5.99.8 Full crystallographic information is available from OCA.
Reference
Structure and kinetics of monofunctional proline dehydrogenase from Thermus thermophilus., White TA, Krishnan N, Becker DF, Tanner JJ, J Biol Chem. 2007 May 11;282(19):14316-27. Epub 2007 Mar 7. PMID:17344208
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