Purification and identification of 91-kDa neutrophil gelatinase. Release by the activating peptide interleukin-8

Eur J Biochem. 1991 Jun 1;198(2):391-8. doi: 10.1111/j.1432-1033.1991.tb16027.x.


Human neutrophils were found to release a 91-kDa gelatinase that is serologically related to tumor-derived gelatinolytic enzymes, as evidenced by immunoprecipitation. In order to identify the neutrophil gelatinase, the activity in conditioned medium from human neutrophil suspensions was purified by affinity chromatography on a gelatin substrate. The 91-kDa active enzyme was further separated from other stainable protein bands by classical SDS PAGE and blotting to a solid support. Amino-terminal sequence analysis of blotted proteins showed that the 91-kDa enzyme is a truncated form of tumor-derived 92-kDa gelatinase (type IV collagenase), lacking eight residues at the NH2-terminus. Sequence analysis of enzymatically inactive cleavage products of this neutrophil gelatinase demonstrated that the gelatin-binding part of the molecule is restricted to the amino-terminal third. Exocytosis of gelatinase-containing granules from neutrophils occurred spontaneously within 6 h after neutrophil plating. When the cells were triggered with the phorbol ester phorbol 12-myristate 13-acetate, a strong secretagogue, rapid gelatinase release was observed. When granulocytes were stimulated with the neutrophil-activating peptide interleukin-8, maximal exocytosis occurred within 1 h. The almost immediate release of neutrophil gelatinase after stimulation of the cells with a chemotactic factor might play a key role in remodeling of the extracellular matrix during granulocyte movement in response to chemotactic stimuli.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cell Line
  • Chromatography, Affinity
  • Electrophoresis, Polyacrylamide Gel
  • Gelatinases
  • Humans
  • Interleukin-1 / pharmacology
  • Interleukin-8 / pharmacology*
  • Molecular Sequence Data
  • Molecular Weight
  • Neutrophils / drug effects
  • Neutrophils / enzymology*
  • Pepsin A / blood*
  • Pepsin A / genetics
  • Pepsin A / isolation & purification
  • Pepsin A / metabolism
  • Peptide Fragments / isolation & purification
  • Sequence Homology, Nucleic Acid
  • Tetradecanoylphorbol Acetate / pharmacology


  • Interleukin-1
  • Interleukin-8
  • Peptide Fragments
  • Pepsin A
  • Gelatinases
  • Tetradecanoylphorbol Acetate

Associated data

  • GENBANK/P14780