1r49

From Proteopedia

proteopedia linkproteopedia link

spinqualitylabelsall models 1r49, resolution 3.13Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image

Human topoisomerase I (Topo70) double mutant K532R/Y723F

Contents 1 Overview 2 Disease 3 About this Structure 4 Reference

Overview

Based on co-crystal structures of human topoisomerase I with bound DNA, Lys(532) makes a minor groove contact with the strongly preferred, thymidine residue at the site of covalent attachment (-1 position)., Replacement of Lys(532) with either arginine or alanine has essentially no, effect on the sequence preference of the enzyme, indicating that this, interaction is not required for the preference for a T at the -1 position., Although both the cleavage and religation activities of the K532R mutant, enzyme are reduced, cleavage is reduced to a greater extent than, religation. The reverse is true for the K532A mutant enzyme with, religation so impaired that the nicked intermediate accumulates during, plasmid relaxation assays. Consistent with the shift in the cleavage, religation equilibrium toward cleavage for the K532A mutant enzyme, expression of the mutant enzyme in Saccharomyces cerevisiae is cytotoxic, and thus this mutant enzyme mimics the effects of the anticancer drug, camptothecin. Cleavage assays with the mutant enzymes using an, oligonucleotide containing a 5'-bridging phosphorothiolate indicate that, Lys(532) functions as a general acid during cleavage to protonate the, leaving 5'-oxygen. It is possible that the contact with the -1 base is, important during catalysis to provide positional rigidity to the active, site. The corresponding residues in the vaccinia virus topoisomerase and, the tyrosine recombinases may have similar critical roles in catalysis.

Disease

Known disease associated with this structure: DNA topoisomerase I, camptothecin-resistant OMIM:[126420]

About this Structure

1R49 is a Single protein structure of sequence from Homo sapiens. Active as DNA topoisomerase, with EC number 5.99.1.2 Full crystallographic information is available from OCA.

Reference

The role of lysine 532 in the catalytic mechanism of human topoisomerase I., Interthal H, Quigley PM, Hol WG, Champoux JJ, J Biol Chem. 2004 Jan 23;279(4):2984-92. Epub 2003 Oct 31. PMID:14594810

Page seeded by OCA on Mon Nov 12 18:59:53 2007