Summer Science Program in Biochemistry - Characterization of the Cdc14 phosphatase homolog from Rhizoctonia solani

Listed in Datasets

By Joyce An1, Caroline Katz, Tudor O Sirbu1, Alex Liu1, Ayush Pandit1, Kathryn A Tso, Mark C Hall2

1. Summer Science Program 2. Purdue University

In SSP Biochemistry, high school participants conduct group research projects to characterize the Cdc14 phosphatase homolog from a phytopathogenic fungus and explore its potential as a novel antifungal target to protect crops from disease

Additional materials available

Version 1.0 - published on 26 Aug 2020 doi:10.4231/DCN9-HC05 - cite this Archived on 26 Aug 2020

Licensed under CC0 1.0 Universal

Gr6_2017_R.solani_Modelimage.jpg Gr6_2017_R.solani_PepBind_image.jpg Gr6_2017_R.solani_PepBind_map.jpg

Description

The Research Project in SSP Biochemistry

Cdc14 enzymes have been reported to be required for normal plant infection by several plant pathogen species. In the SSP Biochemistry program, research groups of 3 participants each purify and characterize the Cdc14 homolog encoded by a different plant fungal pathogen species. Each group is initially given the gene sequence for the putative Cdc14 homolog from a phytopathogenic fungus. Their overall goal is to determine if this protein would be a suitable target for development of a novel fungicide to protect crops from pathogens. They first use bioinformatic tools to generate a hypothesis about the function of the protein encoded by their gene sequence. Then, they use a recombinant plasmid to overexpress their protein in E. coli and purify it using nickel affinity chromatography. They characterize steady-state kinetic properties and substrate specificity of their enzymes and compare the results to previously characterized Cdc14 from the model fungal species Saccharomyces cerevisiae (ScCdc14), for which structural information is available. They then design an assay to test inhibition of their enzyme by a series of compounds originally identified as in vitro inhibitors of ScCdc14. They choose the best one, perform computational docking to predict how it might bind to the homology-based structural model of their enzyme’s active site that they previously generated, and finally hypothesize about ways to optimize their inhibitor structure to improve binding and specificity by taking advantage of known features of Cdc14 substrates. In the end they conclude whether or not the active site substrate specificity is conserved enough to justify using the ScCdc14 structure as a model for designing inhibitors to their enzyme. In this publication, the biochemical results and the homology based structural modeling of the Cdc14 homolog from Rhizoctonia solani are presented.

Cite this work

Researchers should cite this work as follows:

Tags

The Purdue University Research Repository (PURR) is a university core research facility provided by the Purdue University Libraries and the Office of the Executive Vice President for Research and Partnerships, with support from additional campus partners.