Deciphering the Role of 4HVT from Structure to Function

Deciphering the Role of 4HVT from Structure to Function

Alvin Sherman Library

Proteins are molecules crucial to living organisms, as they drive catalytic reactions, cell communication, and cellular structures. Proteins play an important part in all biological processes, and understanding their structure and function is essential for advancing knowledge of life at the molecular level. The protein data bank (PBD) contains thousands of protein structures but many are not functionally characterized. In collaboration with the Seattle Structural Genomics Center for Infectious Disease, this research focuses on elucidating the function of , 4HVT, a hydrolase derived from Rickettsia typhi. The protein was isolated and analyzed using a combination of biochemical and computational techniques to better understand its function. Bioinformatic software tools such as BLAST, SPRITE, DALI, and InterPro were used to determine homologs of 4HVT and form an initial prediction about its function. Homologs are essential to narrowing down hypothesized functions by comparing amino acid sequences, the 3D structure, active sites, and family domains. It was found that four residues of the 4HVT active site are 682 Histidine, 568 Glycine, 566 Serine, and 649 Aspartic Acid. Substrates predicted to interact best with the active sites were determined and confirmed through molecular docking. To better understand the enzyme function, wet lab techniques such as cell lysis, affinity chromatography, protein concentration, SDS-page, and enzyme activity assays were performed. Based on the results of the procedures, the concluded hypothesized function of 4HVT is post-proline peptide cleaving hydrolase EC 3.4.21.26.