1otx
From Proteopedia
(New page: 200px<br /><applet load="1otx" size="450" color="white" frame="true" align="right" spinBox="true" caption="1otx, resolution 2.70Å" /> '''Purine Nucleoside Ph...) |
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| - | [[Image:1otx.gif|left|200px]]<br /><applet load="1otx" size=" | + | [[Image:1otx.gif|left|200px]]<br /><applet load="1otx" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1otx, resolution 2.70Å" /> | caption="1otx, resolution 2.70Å" /> | ||
'''Purine Nucleoside Phosphorylase M64V mutant'''<br /> | '''Purine Nucleoside Phosphorylase M64V mutant'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Activation of prodrugs by Escherichia coli purine nucleoside phosphorylase | + | Activation of prodrugs by Escherichia coli purine nucleoside phosphorylase (PNP) provides a method for selectively killing tumor cells expressing a transfected PNP gene. This gene therapy approach requires matching a prodrug and a known enzymatic activity present only in tumor cells. The specificity of the method relies on avoiding prodrug cleavage by enzymes already present in the host cells or the intestinal flora. Using crystallographic and computer modeling methods as guides, we have redesigned E. coli PNP to cleave new prodrug substrates more efficiently than does the wild-type enzyme. In particular, the M64V PNP mutant cleaves 9-(6-deoxy-alpha-L-talofuranosyl)-6-methylpurine with a kcat/Km over 100 times greater than for native E. coli PNP. In a xenograft tumor experiment, this compound caused regression of tumors expressing the M64V PNP gene. |
==About this Structure== | ==About this Structure== | ||
| - | 1OTX is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with PO4 as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Purine-nucleoside_phosphorylase Purine-nucleoside phosphorylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.2.1 2.4.2.1] Full crystallographic information is available from [http:// | + | 1OTX is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with <scene name='pdbligand=PO4:'>PO4</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Purine-nucleoside_phosphorylase Purine-nucleoside phosphorylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.2.1 2.4.2.1] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1OTX OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Purine-nucleoside phosphorylase]] | [[Category: Purine-nucleoside phosphorylase]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Allan, P | + | [[Category: Allan, P W.]] |
[[Category: Anand, R.]] | [[Category: Anand, R.]] | ||
| - | [[Category: Bennett, E | + | [[Category: Bennett, E M.]] |
| - | [[Category: Ealick, S | + | [[Category: Ealick, S E.]] |
| - | [[Category: Hassan, A | + | [[Category: Hassan, A E.]] |
| - | [[Category: McPherson, D | + | [[Category: McPherson, D T.]] |
| - | [[Category: Parker, W | + | [[Category: Parker, W B.]] |
| - | [[Category: Secrist, J | + | [[Category: Secrist, J A.]] |
| - | [[Category: Sorscher, E | + | [[Category: Sorscher, E J.]] |
[[Category: PO4]] | [[Category: PO4]] | ||
[[Category: empty]] | [[Category: empty]] | ||
[[Category: pnp]] | [[Category: pnp]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:21:31 2008'' |
Revision as of 12:21, 21 February 2008
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Purine Nucleoside Phosphorylase M64V mutant
Overview
Activation of prodrugs by Escherichia coli purine nucleoside phosphorylase (PNP) provides a method for selectively killing tumor cells expressing a transfected PNP gene. This gene therapy approach requires matching a prodrug and a known enzymatic activity present only in tumor cells. The specificity of the method relies on avoiding prodrug cleavage by enzymes already present in the host cells or the intestinal flora. Using crystallographic and computer modeling methods as guides, we have redesigned E. coli PNP to cleave new prodrug substrates more efficiently than does the wild-type enzyme. In particular, the M64V PNP mutant cleaves 9-(6-deoxy-alpha-L-talofuranosyl)-6-methylpurine with a kcat/Km over 100 times greater than for native E. coli PNP. In a xenograft tumor experiment, this compound caused regression of tumors expressing the M64V PNP gene.
About this Structure
1OTX is a Single protein structure of sequence from Escherichia coli with as ligand. Active as Purine-nucleoside phosphorylase, with EC number 2.4.2.1 Full crystallographic information is available from OCA.
Reference
Designer gene therapy using an Escherichia coli purine nucleoside phosphorylase/prodrug system., Bennett EM, Anand R, Allan PW, Hassan AE, Hong JS, Levasseur DN, McPherson DT, Parker WB, Secrist JA 3rd, Sorscher EJ, Townes TM, Waud WR, Ealick SE, Chem Biol. 2003 Dec;10(12):1173-81. PMID:14700625
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