title: Interactions between a viral protease and cystatins.
abstract: The interactions of two cystatins with a viral cysteine protease were studied using several types of assays. Complex formation between the protease and inhibitor was directly demonstrated using a gel retardation assay. It was also shown that the formation of enzyme inhibitor complexes could occur after first binding either the enzyme or the inhibitor to filter paper, and ultimately decorating the complex with antibody and radio-labelled protein A or by preparing one of the protein ligands with an internal radiolabel. The procedure can be adapted to provide a method for screening expression libraries for protease or inhibitor genes. The inhibition of a cysteine protease by a cystatin was shown not to directly involve binding to the active site thiol of the enzyme, but rather to be the result of a steric block in the active site region which prevents large affinity labels and protein substrates from reaching the active site.
authors.affiliation: Central Research and Development Department, DuPont Company, Experimental Station, Wilmington.
authors.forename: B
authors.initials: B
authors.lastname: Korant
authors.forename: T
authors.initials: T
authors.lastname: Towatari
authors.forename: M
authors.initials: M
authors.lastname: Kelley
authors.forename: J
authors.initials: J
authors.lastname: Brzin
authors.forename: B
authors.initials: B
authors.lastname: Lenarcic
authors.forename: V
authors.initials: V
authors.lastname: Turk
mesh_terms: D000818:Animals; D000998:Antiviral Agents; D002645:Chickens; D003001:Cloning, Molecular; D007457:Iodine Radioisotopes; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D013462:Sulfur Radioisotopes; D014361:Trypsin Inhibitors; D014780:Viruses / Q000201:enzymology*
chemical_list: D000998:Antiviral Agents; D007457:Iodine Radioisotopes; D011480:Protease Inhibitors; D013462:Sulfur Radioisotopes; D014361:Trypsin Inhibitors; D010447:Peptide Hydrolases