BASIL2023GVP30646

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Abstract

The Protein Data Bank (PDB) and UniProt contain a wealth of protein structures, many of which do not have a known function. One such protein is P30646, an uncharacterized sugar kinase. Sugar kinases play a role in a variety of metabolic processes within the body, so understanding their functions is of great importance. Our work aimed to further characterize P30646 through the identification of a substrate for this enzyme. Utilizing materials Biochemistry Authentic Scientific Inquiry Laboratory (BASIL) project, our group combined in silico and in vitro data for a full analysis of P30646. Computation tools including BLAST, DALI, SPRITE, and InterPro allowed our group to compare the structural and sequential similarities between P30646 and proteins with known functions. In silico analysis indicated that P30646 had xylulokinase activity, and xylulose was tested as a substrate in vitro using a coupled kinase assay. The kinase assay indicated that P30646 does have strong enzymatic activity for xylulose. The combination of our in silico and in vitro results provide a strong indication that P30646 is a sugar kinase with affinity for xylulose breakdown.

Computational Tools

Laboratory Evidence

Conclusions

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