Start Date
23-4-2025 2:15 PM
Location
3rd floor - Library
Abstract
The mission of the Seattle Structural Genomics Center for Infectious Disease (SSGCID) is to determine the 3D structure of various proteins with important roles in human disease. Many of the proteins they work with have an annotated function but little or no physical characterization. SSGCID partners with other researchers to provide physical characterization of the proteins they work with to support the publication of their structures. From the available structures in the SSGCID database, we chose a probable Blasticidin S deaminase from Coccidioides immitis (PDB ID: 3OJ6) for further characterization. We used several bioinformatic tools to confirm the annotation of this protein, including SPRITE, InterPro, and Dali. These tools gave us confidence in the annotation of this protein as a Blasticidin S deaminase. In the active site, there was a bound zinc ion and glutamate, which acts as a base to activate water as a nucleophile in the reaction; these are key components of a deaminase enzyme. Further physical characterization of this enzyme included determination of its molecular weight by SDS-PAGE and quaternary structure by analytical gel filtration. The enzymatic activity of the protein was determined spectrophotometrically by observing the loss of Blasticidin S. These experiments confirm that 3OJ6 is a Blasticidin S deaminase.
Characterization of a Proposed Blasticidin S Deaminase
3rd floor - Library
The mission of the Seattle Structural Genomics Center for Infectious Disease (SSGCID) is to determine the 3D structure of various proteins with important roles in human disease. Many of the proteins they work with have an annotated function but little or no physical characterization. SSGCID partners with other researchers to provide physical characterization of the proteins they work with to support the publication of their structures. From the available structures in the SSGCID database, we chose a probable Blasticidin S deaminase from Coccidioides immitis (PDB ID: 3OJ6) for further characterization. We used several bioinformatic tools to confirm the annotation of this protein, including SPRITE, InterPro, and Dali. These tools gave us confidence in the annotation of this protein as a Blasticidin S deaminase. In the active site, there was a bound zinc ion and glutamate, which acts as a base to activate water as a nucleophile in the reaction; these are key components of a deaminase enzyme. Further physical characterization of this enzyme included determination of its molecular weight by SDS-PAGE and quaternary structure by analytical gel filtration. The enzymatic activity of the protein was determined spectrophotometrically by observing the loss of Blasticidin S. These experiments confirm that 3OJ6 is a Blasticidin S deaminase.
