title: A new purification procedure of human kidney cathepsin H, its properties and kinetic data.
abstract: A purification procedure of cathepsin H from human kidney is presented. It includes gel filtration, ion exchange chromatography, and covalent chromatography on thiol Sepharose as an essential step. Purified cathepsin H emerges in an isoelectric focusing gel at pH 6.1 and 6.3. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate shows a molecular mass of about 28 kDa. Less than 20% of the enzyme preparation can be separated into a heavy (24 kDa) and a light chain (4 kDa) after reduction and gel filtration on Sephacryl S-200. The partial amino-acid sequence of human cathepsin H shows its close similarity to rat cathepsin H. Inhibition constants (Ki) of cathepsins H and B with chicken cystatin, two forms of human stefin A, human stefin B, and two forms of human cystatin C are in the range of 10(-9) to 10(-11)M.
authors.affiliation: Odsek za biokemijo, Institut J. Stefan, Univerza E. Kardelja, Ljubljana, Jugoslavia.
authors.forename: T
authors.initials: T
authors.lastname: Popović
authors.forename: J
authors.initials: J
authors.lastname: Brzin
authors.forename: J
authors.initials: J
authors.lastname: Kos
authors.forename: B
authors.initials: B
authors.lastname: Lenarcic
authors.forename: W
authors.initials: W
authors.lastname: Machleidt
authors.forename: A
authors.initials: A
authors.lastname: Ritonja
authors.forename: K
authors.initials: K
authors.lastname: Hanada
authors.forename: V
authors.initials: V
authors.lastname: Turk
mesh_terms: D000595:Amino Acid Sequence; D002401:Cathepsin B / Q000037:antagonists & inhibitors; D056655:Cathepsin H; D002403:Cathepsins / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D003546:Cysteine Endopeptidases; D015853:Cysteine Proteinase Inhibitors; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007525:Isoelectric Focusing; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008969:Molecular Sequence Data
chemical_list: D015853:Cysteine Proteinase Inhibitors; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; C534126:CTSH protein, human; D056655:Cathepsin H; C534128:Ctsh protein, rat