Human leukocyte granule elastase: rapid isolation and characterization

Biochemistry. 1976 Feb 24;15(4):836-41. doi: 10.1021/bi00649a017.

Abstract

Human granulocytic elastases have been purified by a two-step procedure involving affinity chromatography of crude extracts of leukocytic granules on Sepharose-Trasylol, followed by ion-exchange chromatography on CM-cellulose to resolve the isoelastases. All of these enzymes were found to be glycoproteins with the carbohydrate content of the major form being composed essentially of only neutral sugars. The molecular weight of this form was found to be near 30 000 daltons with the other forms being slightly higher. Preliminary structural analyses indicate that all of the elastase isozymes have identical NH2-terminal sequences suggesting that the differences in mobility of the four proteins are not due to different degrees of activation from a common zymogen but, more likely, from minor changes in carbohydrate content. Human granulocytic elastases are less active on ligament elastin than porcine pancreatic elastase, but both are inhibited by synthetic elastase active-site directed low molecular weight compounds (Tuhy, P. M., and Powers, J. C. (1975), FEBS Lett. 50, 359) as well as by plasma alpha-1-proteinase inhibitor (formerly called alpha-1-antitrypsin). In the latter case a stable complex with mol wt of 78 000 daltons is formed indicating the formation of a 1:1 complex.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Binding Sites
  • Carbohydrates / analysis
  • Chromatography, Affinity
  • Chromatography, Ion Exchange
  • Cytoplasmic Granules / enzymology*
  • Granulocytes / enzymology*
  • Humans
  • Isoenzymes / analysis*
  • Leukocytes / enzymology*
  • Molecular Weight
  • Pancreatic Elastase / analysis*
  • Pancreatic Elastase / antagonists & inhibitors
  • alpha 1-Antitrypsin / pharmacology

Substances

  • Amino Acids
  • Carbohydrates
  • Isoenzymes
  • alpha 1-Antitrypsin
  • Pancreatic Elastase