2jed
From Proteopedia
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caption="2jed, resolution 2.32Å" /> | caption="2jed, resolution 2.32Å" /> | ||
'''THE CRYSTAL STRUCTURE OF THE KINASE DOMAIN OF THE PROTEIN KINASE C THETA IN COMPLEX WITH NVP-XAA228 AT 2.32A RESOLUTION.'''<br /> | '''THE CRYSTAL STRUCTURE OF THE KINASE DOMAIN OF THE PROTEIN KINASE C THETA IN COMPLEX WITH NVP-XAA228 AT 2.32A RESOLUTION.'''<br /> | ||
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| + | ==Overview== | ||
| + | As exemplified by three cases, we show that the addition of a small molecular weight inhibitor to the culture of Baculovirus-infected insect cells can dramatically improve the expression of a recombinant kinase. The expression of the tyrosine kinase KDR was sevenfold higher and mainly in a soluble form, when the KDR inhibitor PTK/ZK was added to the culture at the time of Baculovirus infection. The expression of the catalytic domain of the serine/threonine kinase PKCtheta, which is otherwise not possible with the Baculovirus expression system, was expressed mainly soluble at 120mg/L by the addition of the PKC inhibitor BIM XI to the culture of Baculovirus-infected insect cells. For Abl kinase, the expression could also be significantly increased by the addition of the Abl kinase inhibitor STI571 to the culture. For all three kinases, this method had previously been applied by us for the improved production of kinase/inhibitor complex protein, leading to the co-crystal structures. It is shown here at the cases KDR-PTK/ZK and PKCtheta-BIM XI, that the stimulatory effect of an inhibitor on kinase expression is applicable under many culture conditions. The presented method represents a valuable tool to obtain structural knowledge on kinase-inhibitor complexes. | ||
==About this Structure== | ==About this Structure== | ||
2JED is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=LG8:'>LG8</scene> and <scene name='pdbligand=MPD:'>MPD</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Protein_kinase_C Protein kinase C], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.13 2.7.11.13] Known structural/functional Sites: <scene name='pdbsite=AC1:Binding+Site+For+Residue+Lg8+A+1701'>AC1</scene>, <scene name='pdbsite=AC2:Binding+Site+For+Residue+Lg8+B+1712'>AC2</scene> and <scene name='pdbsite=AC3:Binding+Site+For+Residue+Mpd+B+1713'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2JED OCA]. | 2JED is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=LG8:'>LG8</scene> and <scene name='pdbligand=MPD:'>MPD</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Protein_kinase_C Protein kinase C], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.11.13 2.7.11.13] Known structural/functional Sites: <scene name='pdbsite=AC1:Binding+Site+For+Residue+Lg8+A+1701'>AC1</scene>, <scene name='pdbsite=AC2:Binding+Site+For+Residue+Lg8+B+1712'>AC2</scene> and <scene name='pdbsite=AC3:Binding+Site+For+Residue+Mpd+B+1713'>AC3</scene>. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2JED OCA]. | ||
| + | |||
| + | ==Reference== | ||
| + | Improved expression of kinases in Baculovirus-infected insect cells upon addition of specific kinase inhibitors to the culture helpful for structural studies., Strauss A, Fendrich G, Horisberger MA, Liebetanz J, Meyhack B, Schlaeppi JM, Schmitz R, Protein Expr Purif. 2007 Dec;56(2):167-76. Epub 2007 Jul 10. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17720535 17720535] | ||
[[Category: Homo sapiens]] | [[Category: Homo sapiens]] | ||
[[Category: Protein kinase C]] | [[Category: Protein kinase C]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Berner, A.]] | [[Category: Berner, A.]] | ||
| - | [[Category: Bitsch]] | + | [[Category: Bitsch, F.]] |
[[Category: Buelens, F.]] | [[Category: Buelens, F.]] | ||
[[Category: Depersin, H.]] | [[Category: Depersin, H.]] | ||
| - | [[Category: F]] | ||
[[Category: Geiser, M.]] | [[Category: Geiser, M.]] | ||
[[Category: Graff, P.]] | [[Category: Graff, P.]] | ||
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[[Category: Rahuel, J.]] | [[Category: Rahuel, J.]] | ||
[[Category: Rummel, G.]] | [[Category: Rummel, G.]] | ||
| - | [[Category: Schlaeppi, J | + | [[Category: Schlaeppi, J M.]] |
[[Category: Schmitz, R.]] | [[Category: Schmitz, R.]] | ||
[[Category: Stark, W.]] | [[Category: Stark, W.]] | ||
| Line 40: | Line 45: | ||
[[Category: zinc-finger]] | [[Category: zinc-finger]] | ||
| - | ''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 18:02:20 2008'' |
Revision as of 16:02, 21 February 2008
|
THE CRYSTAL STRUCTURE OF THE KINASE DOMAIN OF THE PROTEIN KINASE C THETA IN COMPLEX WITH NVP-XAA228 AT 2.32A RESOLUTION.
Overview
As exemplified by three cases, we show that the addition of a small molecular weight inhibitor to the culture of Baculovirus-infected insect cells can dramatically improve the expression of a recombinant kinase. The expression of the tyrosine kinase KDR was sevenfold higher and mainly in a soluble form, when the KDR inhibitor PTK/ZK was added to the culture at the time of Baculovirus infection. The expression of the catalytic domain of the serine/threonine kinase PKCtheta, which is otherwise not possible with the Baculovirus expression system, was expressed mainly soluble at 120mg/L by the addition of the PKC inhibitor BIM XI to the culture of Baculovirus-infected insect cells. For Abl kinase, the expression could also be significantly increased by the addition of the Abl kinase inhibitor STI571 to the culture. For all three kinases, this method had previously been applied by us for the improved production of kinase/inhibitor complex protein, leading to the co-crystal structures. It is shown here at the cases KDR-PTK/ZK and PKCtheta-BIM XI, that the stimulatory effect of an inhibitor on kinase expression is applicable under many culture conditions. The presented method represents a valuable tool to obtain structural knowledge on kinase-inhibitor complexes.
About this Structure
2JED is a Single protein structure of sequence from Homo sapiens with and as ligands. Active as Protein kinase C, with EC number 2.7.11.13 Known structural/functional Sites: , and . Full crystallographic information is available from OCA.
Reference
Improved expression of kinases in Baculovirus-infected insect cells upon addition of specific kinase inhibitors to the culture helpful for structural studies., Strauss A, Fendrich G, Horisberger MA, Liebetanz J, Meyhack B, Schlaeppi JM, Schmitz R, Protein Expr Purif. 2007 Dec;56(2):167-76. Epub 2007 Jul 10. PMID:17720535
Page seeded by OCA on Thu Feb 21 18:02:20 2008
Categories: Homo sapiens | Protein kinase C | Single protein | Berner, A. | Bitsch, F. | Buelens, F. | Depersin, H. | Geiser, M. | Graff, P. | Knecht, R. | Rahuel, J. | Rummel, G. | Schlaeppi, J M. | Schmitz, R. | Stark, W. | Strauss, A. | Wagner, J. | LG8 | MPD | Alternative splicing | Atp-binding | Kinase | Magnesium | Metal-binding | Nucleotide-binding | Phorbol-ester binding | Phosphorylation | Pkc theta | Polymorphism | Serine/threonine-protein kinase | Transferase | Zinc | Zinc-finger
