A Multilayered Approach to Predict Metal-Binding Sites in the Pneumococcal Phosphoglucomutase Protein
Additional Funding Sources
The project was funded by the Ronald E. McNair Post Baccalaureate Achievement Program at Idaho State University through the Department of Education under Award No. P217A170169.
Abstract
Phosphoglucomutase (Pgm) is an important enzyme that functions to bridge sugar metabolism to capsular polysaccharide formation in Streptococcus pneumoniae. Preliminary data using a mag-fura-2 metal-binding competition assay reveals that pneumococcal Pgm binds 2 metal ions per protomer, despite literature showing a single metal-binding site for other characterized Pgm family members. In this study, we focus on identifying the second metal-binding site in the pneumococcal Pgm. I-TASSER was used to generate a structural model of the pneumococcal Pgm. A multi-protein sequence alignment comparing pneumococcal Pgm and other bacterial Pgm proteins using multiple bioinformatics programs validated the known highly conserved metal-binding motif DXDXDR, as well as several other potential metal-binding motifs. We share information here on two of these secondary predicted metal-binding motifs. One motif is found downstream of the Pgm active site and the other is located near the C-terminus of the protein. These preliminary data set the foundation for further biochemical and phenotypical analysis of mutant Pgm protein.
A Multilayered Approach to Predict Metal-Binding Sites in the Pneumococcal Phosphoglucomutase Protein
Phosphoglucomutase (Pgm) is an important enzyme that functions to bridge sugar metabolism to capsular polysaccharide formation in Streptococcus pneumoniae. Preliminary data using a mag-fura-2 metal-binding competition assay reveals that pneumococcal Pgm binds 2 metal ions per protomer, despite literature showing a single metal-binding site for other characterized Pgm family members. In this study, we focus on identifying the second metal-binding site in the pneumococcal Pgm. I-TASSER was used to generate a structural model of the pneumococcal Pgm. A multi-protein sequence alignment comparing pneumococcal Pgm and other bacterial Pgm proteins using multiple bioinformatics programs validated the known highly conserved metal-binding motif DXDXDR, as well as several other potential metal-binding motifs. We share information here on two of these secondary predicted metal-binding motifs. One motif is found downstream of the Pgm active site and the other is located near the C-terminus of the protein. These preliminary data set the foundation for further biochemical and phenotypical analysis of mutant Pgm protein.