36. Random Peptide Library Based on a Spider Neurotoxin, and Utilization of the Library in in vitro Evolution Directed to GPCR Ligands

High Proteolytic Resistance of Spider-Derived Inhibitor Cystine Knots

“…The original function can be optimized and/or new functionalities can be grafted to existing ICK scaffolds and such engineered ICKs are reported to be applied to HIV vaccine [ 46 ] and tumor targeting [ 39 , 47 ]. We have also prepared a random peptide library using GTx1-15 as a scaffold to tune loop sequences with PERISS (intraperiplasm secretion and selection) method, a kind of directed evolution in which a target protein and the interacting peptide are expressed in E. coli inner membrane and periplasmic space, respectively [ 48 ], and we will soon be able to report what we obtain with GTx1-15 scaffold and its pharmacokinetic data.…”

The application of the Escherichia coli giant spheroplast for drug screening with automated planar patch clamp system

“…Again, the most important advantage of all is that the direct use of bacterial spheroplasts can skip painstaking steps needed to set up the eukaryotic system for electrophysiological analysis. Now we are refining a novel technique named PERRIS (intra periplasm secretion and selection) method, a kind of in vitro directed evolution in which a target protein and the interacting peptide are expressed in E. coli inner membrane and periplasmic space, respectively, to search and optimize interacting peptides [12] . In this process, tremendous number of recombinant E. coli would be produced and we needed a fast and inexpensive way of primary screening.…”