Photochemical protease: Site-specific photocleavage of hen egg lysozyme and bovine serum albumin

Abstract

Site-specific photocleavage of hen egg lysozyme and bovine serum albumin (

Chiral protein scissors: High enantiomeric selectivity for binding and its effect on protein photocleavage efficiency and specificity

“…To separate the reaction mixture containing BSA, 8% polyacrylamide (separating) gel was used while the lysozyme reaction mixture was separated by using a 12% polyacrylamide gel. The gel was electrophoresed by applying 60 V until the dye passed into the separating gel, and then the voltage was increased to 100 V. The gels were run for 2 h, stained, and destained as described earlier (19).…”

“…These residues are located in the interior of BSA, near the fatty acid-binding site, in domain II, subdomain C. In contrast, lysozyme was cleaved at a site exposed to the solvent, between Trp-108 and Val-109, away from the catalytic site. In addition to cleaving at a single site, the photoreaction did not alter the enzymatic activity of lysozyme, indicating the gentle nature of the cleavage chemistry toward the sensitive residues at the catalytic site (19). Control over the location of the photocleavage site was demonstrated when additional residues were inserted between the pyrenyl chromophore and phenylalanine of 4(1-pyrene)-3-butyroyl-phenylalanine amide (Py-Phe) (20).…”

“…Site-specific photocleavage of proteins with high efficiency by using probes based on a pyrenyl chromophore coupled to an asymmetric center was reported recently (4)(5)(6)(7)(8)19). By linking a photoreactive chromophore to natural amino acids or short peptides, protein photocleavage at specific sites was demonstrated.…”

“…By linking a photoreactive chromophore to natural amino acids or short peptides, protein photocleavage at specific sites was demonstrated. Covalent linking of 4(1-pyrenyl)butyric acid to the N terminus of L-phenylalanine, for example, led to a photoprobe (Py-L-Phe, Scheme 1) that cleaves BSA or lysozyme at a single site with high quantum efficiency (0.25-0.003) (19). The newly produced photofragments are amenable to sequencing, and BSA was cleaved by Py-L-Phe between Leu-346 and Arg-347.…”

Study on the interaction of Prodigiosin with bovine serum albumin by spectroscopic methods

“…Serum albumin is the major protein constituent of blood plasma which facilitates the disposition and transportation of various exogenous and endogenous ligands to the specific targets [16]. Bovine serum Scheme 1.…”