After 20+ years of determining atomic level structures of proteins in three dimensions, we are going properly 3D. Our Prusa i3 MKS3+ has arrived!
22
Feb
2023
Month: February 2023
11
Feb
2023
Evolution of protease activation and specificity via alpha-2-macroglobulin-mediated covalent capture
Nature Communications 14:768 (2023)
DOI: 10.1038/s41467-023-36099-7
Pubmed: 36765057
PDB coordinates:6YV5 (3D view ), 6YV6 (3D view )
Abstract
Tailoring of the activity and specificity of proteases is critical for their utility across industrial, medical and research purposes. However, engineering or evolving protease catalysts is challenging and often labour intensive. Here, we describe a generic method to accelerate this process based on yeast display. We introduce the protease selection system A2Mcap that covalently captures protease catalysts by repurposed alpha-2-macroglobulin (A2Ms). To demonstrate the utility of A2Mcap for protease engineering we exemplify the directed activity and specificity evolution of six serine proteases. This resulted in a variant of Staphylococcus aureus serin-protease-like (Spl) protease SplB, an enzyme used for recombinant protein processing, that no longer requires activation by N-terminal signal peptide removal. Continue reading →
01
Feb
2023
One of those weeks
HPLC plays up with random UV peaks, fraction collector tries to fail its calibration, centrifuge bottles crack with half of mammalian expression culture lost, fluorescent detector cuts off randomly.
And it’s only a Wednesday.
Panton Arms, here we come.