2VZ0

Pteridine Reductase 1 (PTR1) from Trypanosoma Brucei in complex with NADP and DDD00066641

Summary for 2VZ0

• Entry DOI: 10.2210/pdb2vz0/pdb
• Related: 2C7V
• Descriptor: PTERIDINE REDUCTASE, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, 6-(4-methylphenyl)quinazoline-2,4-diamine, ... (4 entities in total)
• Functional Keywords: oxidoreductase, short-chain dehydrogenase/reductase, trypanosomatids, pteridine reductase
• Biological source: TRYPANOSOMA BRUCEI BRUCEI
• Total number of polymer chains: 4
• Total formula weight: 117968.58

Authors

Robinson, D.A.,Thompson, S.,Sienkiewicz, N.,Fairlamb, A.H. (deposition date: 2008-07-29, release date: 2009-09-22, Last modification date: 2023-12-13)

Primary citation

Shanks, E.J.,Ong, H.B.,Robinson, D.A.,Thompson, S.,Sienkiewicz, N.,Fairlamb, A.H.,Frearson, J.A.
Development and Validation of a Cytochrome C Coupled Assay for Pteridine Reductase 1 and Dihydrofolate Reductase.
Anal.Biochem., 396:194-, 2010

PubMed Abstract: Activity of the pterin- and folate-salvaging enzymes pteridine reductase 1 (PTR1) and dihydrofolate reductase-thymidylate synthetase (DHFR-TS) is commonly measured as a decrease in absorbance at 340 nm, corresponding to oxidation of nicotinamide adenine dinucleotide phosphate (NADPH). Although this assay has been adequate to study the biology of these enzymes, it is not amenable to support any degree of routine inhibitor assessment because its restricted linearity is incompatible with enhanced throughput microtiter plate screening. In this article, we report the development and validation of a nonenzymatically coupled screening assay in which the product of the enzymatic reaction reduces cytochrome c, causing an increase in absorbance at 550 nm. We demonstrate this assay to be robust and accurate, and we describe its utility in supporting a structure-based design, small-molecule inhibitor campaign against Trypanosoma brucei PTR1 and DHFR-TS.

PubMed: 19748480
DOI: 10.1016/J.AB.2009.09.003

Experimental method

X-RAY DIFFRACTION (1.9 Å)