Progelatinase B Forms from Human Neutrophils. Complex Formation of Monomer/Lipocalin with TIMP-1

• Published in: Biological Chemistry Vol. 377; no. 7-8; pp. 529 - 534
• Main Authors: Kolkenbrock, Hansjörg, Hecker-Kia, Adelheid, Orgel, Dagmar, Kinawi, Abdelwahab, Ulbrich, Norbert
• Format: Journal Article
• Language: English
• Published: Berlin, New York Walter de Gruyter, Berlin / New York 01.07.1996
• Subjects: Carrier Proteins - metabolism; Enzyme Activation; Enzyme Precursors - metabolism; Gelatinases - metabolism; Glycoproteins - metabolism; Humans; Matrix Metalloproteinase 3 - metabolism; Matrix Metalloproteinase 9; Matrix Metalloproteinase Inhibitors; Metalloendopeptidases - metabolism; Neutrophils - enzymology; Tissue Inhibitor of Metalloproteinases
• ISSN: 0177-3593; 1431-6730; 1437-4315
• DOI: 10.1515/bchm3.1996.377.7-8.529

The three forms of neutrophil gelatinase B-monomer, homodimer and monomer/lipocalin complex-, were isolated from phorbolester stimulated neutrophil granulocytes by chromatography on gelatin-Sepharose and heparin-Ultrogel. On average, about 50% of the monomer/lipocalin complex was found to be complexed with TIMP-1. After activation with trypsin monomer, homodimer and monomer/lipocalin complex displayed a specific activity of about 2000 mU/mg towards the substrate N-(2,4)-dinitrophenyl-Pro-Gln-Gly-lle-Ala-Gly-Gln-D-Arg, whereas the monomer/lipocalin/TIMP-1 complex could be activated to a specific activity of only 200 mU/mg. The ternary monomer/lipocalin/TIMP-1 complex behaves like the progelatinase A-TIMP-2 complex and the progelatinase B-TIMP-1 complex in that it is an inhibitor for active metalloproteinases (MMPs) and, after activation, a gelatinase with a pronouncedly reduced activity. When the monomer/lipocalin/TIMP-1 complex inhibits an MMP, a quaternary complex monomer/lipocalin/TIMP-1/MMP is generated which after activation shows a sixfold higher proteolytic activity than the active ternary complex.