Evidence for the tissue inhibitor of metalloproteinases-1 (TIMP-1) in human polymorphonuclear leukocytes.
A tissue inhibitor of metalloproteinases (TIMP)-1 was isolated from human polymorphonuclear leukocytes (PMNL) in a complex with latent 95-kDa gelatinase (matrixmetalloproteinase, MMP-9). It was separated from the enzyme by gel filtration in the presence of SDS. Using a competitive ELISA procedure, we determined that 10% of the isolated gelatinase was complexed with TIMP-1. The presence of the inhibitor in isolated PMNL could also be demonstrated by indirect immunofluorescence using a specific antibody against TIMP-1. Cellular mRNA was isolated from PMNL, which were highly purified by separation via formylMet-Leu-Pro-stimulated chemotactic migration in a Boyden chamber. Using reverse-transcription PCR and Northern blotting, TIMP-1 mRNA was shown to be present in PMNL, suggesting that these cells are also capable of synthesizing TIMP-1.
Triebel
S
S
Department of Biochemistry, University of Bielefeld, Germany.
Bläser
J
J
Gote
T
T
Pelz
G
G
Schüren
E
E
Schmitt
M
M
Tschesche
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D018093:Gelatinases / Q000378:metabolism; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger / Q000235:genetics; D019714:Tissue Inhibitor of Metalloproteinases
D006023:Glycoproteins; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D019714:Tissue Inhibitor of Metalloproteinases; D018093:Gelatinases