Identification of a new serine hydrolase
Start Date
2023 2:15 PM
Location
Alter Hall Poster Session 1 - 2nd floor
Abstract
Of the over 10,000 proteins that were crystallized and deposited into the Protein Data Bank during the Protein Structure Initiative, roughly 30% of them remain without a defined function. In this project, the functions of these proteins are assigned using a combination of in silico and wet-lab techniques. Computational tools are used to generate a preliminary hypothetical function for the protein. Following the BASIL curriculum, SPRITE is used to identify an arrangement of amino acids with a known function. BLAST and Pfam are used to compare the sequence of the new protein with known sequences. DALI compares the overall structure of this protein with other proteins of known function.
After generating a reasonable hypothesis of likely protein function, this hypothesis is tested using wet-lab experiments. Expression clones for the protein were obtained from the DNASU repository and the protein was expressed and purified using Ni-affinity chromatography. Docking experiments using SwissDock were used to identify potential substrates for kinetic analysis. Enzymatic activity of the new protein with the substrate identified by docking confirms its function.
Identification of a new serine hydrolase
Alter Hall Poster Session 1 - 2nd floor
Of the over 10,000 proteins that were crystallized and deposited into the Protein Data Bank during the Protein Structure Initiative, roughly 30% of them remain without a defined function. In this project, the functions of these proteins are assigned using a combination of in silico and wet-lab techniques. Computational tools are used to generate a preliminary hypothetical function for the protein. Following the BASIL curriculum, SPRITE is used to identify an arrangement of amino acids with a known function. BLAST and Pfam are used to compare the sequence of the new protein with known sequences. DALI compares the overall structure of this protein with other proteins of known function.
After generating a reasonable hypothesis of likely protein function, this hypothesis is tested using wet-lab experiments. Expression clones for the protein were obtained from the DNASU repository and the protein was expressed and purified using Ni-affinity chromatography. Docking experiments using SwissDock were used to identify potential substrates for kinetic analysis. Enzymatic activity of the new protein with the substrate identified by docking confirms its function.
