Elastase released from human granulocytes stimulated with N-formyl-chemotactic peptide prevents activation of tumor cell prourokinase (pro-uPA).
Proteolytic enzymes released from granulocytes upon stimulation with the chemotactic N-formyl peptide FNLPNTL (in the presence of cytochalasin B) prevented activation of tumor cell single-chain urokinase-type plasminogen activator (pro-uPA) by plasmin. Elastase was identified by the use of eglin C (elastase inhibitor) and a monoclonal antibody to elastase as the functional proteolytic enzyme in granulocyte supernatants. Action of purified granulocyte elastase on pro-uPA generated enzymatically inactive two-chain uPA linked by disulfide bridges which was indistinguishable by SDS-PAGE from plasmin-generated HMW-uPA. The major elastase cleavage site in pro-uPA was located between Ile159 and Ile160. a minor one between Thr165 and Thr166. Elastase cannot substitute for plasmin in the proteolytic activation of pro-uPA to enzymatically active HMW-uPA. However, when pro-uPA was first activated by plasmin to form enzymatically active HMW-uPA, this enzymatic activity was not impaired by subsequent elastase treatment.
Schmitt
M
M
Frauenklinik der Technischen Universität München, FRG.
Kanayama
N
N
Henschen
A
A
Hollrieder
A
A
Hafter
R
R
Gulba
D
D
Jänicke
F
F
Graeff
H
H
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D002630:Chemotactic Factors / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005341:Fibrinolysin / Q000494:pharmacology; D006098:Granulocytes / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000494:pharmacology*; D010196:Pancreatic Elastase / Q000097:blood* / Q000494:pharmacology; D010960:Plasminogen Activators / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015849:Plasminogen Inactivators; D011506:Proteins; D015842:Serine Proteinase Inhibitors; D015843:Serpins; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000596:Amino Acids; D002630:Chemotactic Factors; D009842:Oligopeptides; D015849:Plasminogen Inactivators; D011506:Proteins; D015842:Serine Proteinase Inhibitors; D015843:Serpins; C028656:eglin proteinase inhibitors; C021642:F-chemotactic peptide; D010960:Plasminogen Activators; D010196:Pancreatic Elastase; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator