Figure 6

Protease activity screen using a fluorescence substrate, BZAR. A. The BZAR (Rhodamine 110, bis-(N-CBZ-L-arginine amide), dihydrochloride) is a general substrate of serine proteases. Upon protease cleavage, the nonfluorescent bisamide derivative of rhodamine 110 is first converted to the fluorescent monoamide and then to rhodamine 110, with a further increase in fluorescence. B. Time course measurements of protease cleavage of BZAR. The protease cleavage reactions were montiored by time fluorescence measurements (λ_ex: 485 nm, λ_em: 535 nm) using a GENeo Pro (Tecan). The increase of fluorescence represents protease cleavage reactions. Δ: y3353, putative kinase, ○: trypsin, ◊: y1992: putative carboxypeptidase, and □: SP1780, oligoendopeptidase.