A new purification procedure of human kidney cathepsin H, its properties and kinetic data

Biol Chem Hoppe Seyler. 1988 May:369 Suppl:175-83.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cathepsin B / antagonists & inhibitors
  • Cathepsin H
  • Cathepsins / antagonists & inhibitors
  • Cathepsins / isolation & purification*
  • Cathepsins / metabolism
  • Cysteine Endopeptidases*
  • Cysteine Proteinase Inhibitors
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Isoelectric Focusing
  • Kidney / enzymology*
  • Kinetics
  • Molecular Sequence Data

Substances

  • Cysteine Proteinase Inhibitors
  • Cathepsins
  • Cysteine Endopeptidases
  • Cathepsin B
  • CTSH protein, human
  • Cathepsin H
  • Ctsh protein, rat