Application of biophysical methods for improved protein production and characterization: A case study on an high-temperature requirement A-family bacterial protease.

ABSTRACT: The high-temperature requirement A (HtrA) serine protease family presents an attractive target class for antibacterial therapeutics development. These proteins possess dual protease and chaperone functions and contain numerous binding sites and regulatory loops, displaying diverse oligomerization patterns dependent on substrate type and occupancy. HtrA proteins that are natively purified coelute with contaminating peptides and activating species, shifting oligomerization and protein structure to differently activated populations. Here, a redesigned HtrA production results in cleaner preparations with high yields by overexpressing and purifying target protein from inclusion bodies under denaturing conditions, followed by a high-throughput screen for optimal refolding buffer composition usin

SUBMITTER: Ronzetti M 

PROVIDER: S-EPMC9679970 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature