1ly8
From Proteopedia
(New page: 200px<br /><applet load="1ly8" size="450" color="white" frame="true" align="right" spinBox="true" caption="1ly8, resolution 2.05Å" /> '''The crystal structur...) |
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| - | [[Image:1ly8.jpg|left|200px]]<br /><applet load="1ly8" size=" | + | [[Image:1ly8.jpg|left|200px]]<br /><applet load="1ly8" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1ly8, resolution 2.05Å" /> | caption="1ly8, resolution 2.05Å" /> | ||
'''The crystal structure of a mutant enzyme of Coprinus cinereus peroxidase provides an understanding of its increased thermostability and insight into modelling of protein structures'''<br /> | '''The crystal structure of a mutant enzyme of Coprinus cinereus peroxidase provides an understanding of its increased thermostability and insight into modelling of protein structures'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Seven amino-acid substitutions introduced into the 343 amino-acid-long | + | Seven amino-acid substitutions introduced into the 343 amino-acid-long sequence of Coprinus cinereus peroxidase (CiP) led to a mutant enzyme (TS-rCiP) which is more stable than the native enzyme at higher temperature, pH and hydrogen peroxide concentrations. It is therefore more suitable for industrial applications. A structure determination was conducted on a deglycosylated but still active form of TS-rCiP based on X-ray diffraction data to 2.05 A resolution measured on a crystal cooled to 100 K and refined to R = 0.202 and R(free) = 0.249. The increased stability of the TS-rCiP enzyme can be understood from the structural changes of the TS-rCiP structure revealed by a comparative analysis with other known CiP structures. One of the more significant changes caused by three of the substitutions, I49S, V53A and T121A, is the conversion of a hydrophobic pocket into a hydrophilic pocket with associated changes in the water structure and the hydrogen-bonding interactions. The E239G substitution, which gives rise to increased thermostability at high pH, creates changes in the water structure and in the orientation of a phenylalanine (Phe236) in its vicinity. The three substitutions M166F, M242 and Y242F introduced to increase the oxidative stability do not introduce any structural changes. |
==About this Structure== | ==About this Structure== | ||
| - | 1LY8 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Coprinopsis_cinerea Coprinopsis cinerea] with NAG, BMA, MAN, CA, HEM and GOL as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Peroxidase Peroxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.11.1.7 1.11.1.7] Full crystallographic information is available from [http:// | + | 1LY8 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Coprinopsis_cinerea Coprinopsis cinerea] with <scene name='pdbligand=NAG:'>NAG</scene>, <scene name='pdbligand=BMA:'>BMA</scene>, <scene name='pdbligand=MAN:'>MAN</scene>, <scene name='pdbligand=CA:'>CA</scene>, <scene name='pdbligand=HEM:'>HEM</scene> and <scene name='pdbligand=GOL:'>GOL</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Peroxidase Peroxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.11.1.7 1.11.1.7] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1LY8 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Houborg, K.]] | [[Category: Houborg, K.]] | ||
[[Category: Larsen, S.]] | [[Category: Larsen, S.]] | ||
| - | [[Category: Poulsen, J | + | [[Category: Poulsen, J C.N.]] |
[[Category: Schneider, P.]] | [[Category: Schneider, P.]] | ||
[[Category: Svendsen, A.]] | [[Category: Svendsen, A.]] | ||
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[[Category: thermostability]] | [[Category: thermostability]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:49:35 2008'' |
Revision as of 11:49, 21 February 2008
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The crystal structure of a mutant enzyme of Coprinus cinereus peroxidase provides an understanding of its increased thermostability and insight into modelling of protein structures
Overview
Seven amino-acid substitutions introduced into the 343 amino-acid-long sequence of Coprinus cinereus peroxidase (CiP) led to a mutant enzyme (TS-rCiP) which is more stable than the native enzyme at higher temperature, pH and hydrogen peroxide concentrations. It is therefore more suitable for industrial applications. A structure determination was conducted on a deglycosylated but still active form of TS-rCiP based on X-ray diffraction data to 2.05 A resolution measured on a crystal cooled to 100 K and refined to R = 0.202 and R(free) = 0.249. The increased stability of the TS-rCiP enzyme can be understood from the structural changes of the TS-rCiP structure revealed by a comparative analysis with other known CiP structures. One of the more significant changes caused by three of the substitutions, I49S, V53A and T121A, is the conversion of a hydrophobic pocket into a hydrophilic pocket with associated changes in the water structure and the hydrogen-bonding interactions. The E239G substitution, which gives rise to increased thermostability at high pH, creates changes in the water structure and in the orientation of a phenylalanine (Phe236) in its vicinity. The three substitutions M166F, M242 and Y242F introduced to increase the oxidative stability do not introduce any structural changes.
About this Structure
1LY8 is a Single protein structure of sequence from Coprinopsis cinerea with , , , , and as ligands. Active as Peroxidase, with EC number 1.11.1.7 Full crystallographic information is available from OCA.
Reference
The structure of a mutant enzyme of Coprinus cinereus peroxidase provides an understanding of its increased thermostability., Houborg K, Harris P, Poulsen JC, Schneider P, Svendsen A, Larsen S, Acta Crystallogr D Biol Crystallogr. 2003 Jun;59(Pt 6):997-1003. Epub 2003, May 23. PMID:12777761
Page seeded by OCA on Thu Feb 21 13:49:35 2008
Categories: Coprinopsis cinerea | Peroxidase | Single protein | Harris, P. | Houborg, K. | Larsen, S. | Poulsen, J C.N. | Schneider, P. | Svendsen, A. | BMA | CA | GOL | HEM | MAN | NAG | Coprinus cinereus | Mutant | Thermostability
