Metalloprotease Research
Start Date
August 2025
End Date
August 2025
Location
ALT 303
Abstract
BASIL is a course-based undergraduate research experience that includes using a computer to go through modules that direct students step-by-step through various websites to assign functions to proteins. BASIL is comprised of four main modules: SPRITE, BLAST, Pfam, and DALI/Foldseek. These websites determine protein function by comparing protein sequence and structure to already known proteins located in the PDB. Specifically in this research, students were given a large list of metalloproteases, which are proteins with metals in them, and tasked to find a list of potential candidates that have unknown functions so that in later dates, other students can test their functions in a wet laboratory setting. Using the BASIL modules, a large list of metalloproteases was shortened to a list of 13, then upon further analysis using DNASU and other sources we narrowed it down to a list of 7 potential candidates to be further examined for their functions in a wet lab. Next, 3CBW, an already known BASIL protein, was purified and expressed to understand the procedure needed for the metalloproteases. The protein was then run through kinetics to confirm its function based on past research. Kinetic analysis was run with para-nitrophenyl butyrate as well as AZCL-galactomannan (Megazyme), and it was concluded the 3CBW was able to hydrolyze both substrates. The results of these tests led to the hypothesis that 3CBW is likely a beta-mannosidase.
Metalloprotease Research
ALT 303
BASIL is a course-based undergraduate research experience that includes using a computer to go through modules that direct students step-by-step through various websites to assign functions to proteins. BASIL is comprised of four main modules: SPRITE, BLAST, Pfam, and DALI/Foldseek. These websites determine protein function by comparing protein sequence and structure to already known proteins located in the PDB. Specifically in this research, students were given a large list of metalloproteases, which are proteins with metals in them, and tasked to find a list of potential candidates that have unknown functions so that in later dates, other students can test their functions in a wet laboratory setting. Using the BASIL modules, a large list of metalloproteases was shortened to a list of 13, then upon further analysis using DNASU and other sources we narrowed it down to a list of 7 potential candidates to be further examined for their functions in a wet lab. Next, 3CBW, an already known BASIL protein, was purified and expressed to understand the procedure needed for the metalloproteases. The protein was then run through kinetics to confirm its function based on past research. Kinetic analysis was run with para-nitrophenyl butyrate as well as AZCL-galactomannan (Megazyme), and it was concluded the 3CBW was able to hydrolyze both substrates. The results of these tests led to the hypothesis that 3CBW is likely a beta-mannosidase.
