The axonally secreted serine proteinase inhibitor, neuroserpin, inhibits plasminogen activators and plasmin but not thrombin.
Neuroserpin is an axonally secreted serine proteinase inhibitor that is expressed in neurons during embryogenesis and in the adult nervous system. To identify target proteinases, we used a eucaryotic expression system based on the mouse myeloma cell line J558L and vectors including a promoter from an Ig-kappa-variable region, an Ig-kappa enhancer, and the exon encoding the Ig-kappa constant region (C kappa) and produced recombinant neuroserpin as a wild-type protein or as a fusion protein with C kappa. We investigated the capability of recombinant neuroserpin to form SDS-stable complexes with, and to reduce the amidolytic activity of, a variety of serine proteinases in vitro. Consistent with its primary structure at the reactive site, neuroserpin exhibited inhibitory activity against trypsin-like proteinases. Although neuroserpin bound and inactivated plasminogen activators and plasmin, no interaction was observed with thrombin. A reactive site mutant of neuroserpin neither formed complexes with nor inhibited the amidolytic activity of any of the tested proteinases. Kinetic analysis of the inhibitory activity revealed neuroserpin to be a slow binding inhibitor of plasminogen activators and plasmin. Thus, we postulate that neuroserpin could represent a regulatory element of extracellular proteolytic events in the nervous system mediated by plasminogen activators or plasmin.
Osterwalder
T
T
Institute of Biochemistry, University of Zurich, Switzerland.
Cinelli
P
P
Baici
A
A
Pennella
A
A
Krueger
S R
SR
Schrimpf
S P
SP
Meins
M
M
Sonderegger
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001369:Axons / Q000378:metabolism*; D002642:Chick Embryo; D005341:Fibrinolysin / Q000037:antagonists & inhibitors*; D005822:Genetic Vectors; D006023:Glycoproteins / Q000378:metabolism*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000378:metabolism*; D010960:Plasminogen Activators / Q000037:antagonists & inhibitors*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D015842:Serine Proteinase Inhibitors / Q000378:metabolism*; D015843:Serpins / Q000378:metabolism*; D013917:Thrombin / Q000037:antagonists & inhibitors*; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors; D000097522:Neuroserpin
D005341:Fibrinolysin; D006023:Glycoproteins; D007145:Immunoglobulin kappa-Chains; D009479:Neuropeptides; D010960:Plasminogen Activators; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D015843:Serpins; D013917:Thrombin; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D000097522:Neuroserpin