1OTX

Purine Nucleoside Phosphorylase M64V mutant

Summary for 1OTX

• Entry DOI: 10.2210/pdb1otx/pdb
• Related: 1A90 1ECP 1OTY
• Descriptor: Purine nucleoside phosphorylase, PHOSPHATE ION (3 entities in total)
• Functional Keywords: empty, pnp, transferase
• Biological source: Escherichia coli
• Total number of polymer chains: 3
• Total formula weight: 77740.96

Authors

Ealick, S.E.,Bennett, E.M.,Anand, R.,Secrist, J.A.,Parker, W.B.,Hassan, A.E.,Allan, P.W.,McPherson, D.T.,Sorscher, E.J. (deposition date: 2003-03-23, release date: 2004-02-17, Last modification date: 2023-08-16)

Primary citation

Bennett, E.M.,Anand, R.,Allan, P.W.,Hassan, A.E.,Hong, J.S.,Levasseur, D.N.,McPherson, D.T.,Parker, W.B.,Secrist, J.A.,Sorscher, E.J.,Townes, T.M.,Waud, W.R.,Ealick, S.E.
Designer gene therapy using an Escherichia coli purine nucleoside phosphorylase/prodrug system.
Chem.Biol., 10:1173-1181, 2003

PubMed Abstract: 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.

PubMed: 14700625
DOI: 10.1016/j.chembiol.2003.11.008

Experimental method

X-RAY DIFFRACTION (2.7 Å)