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 a...

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Bibliographic Details
Published in:	The Journal of biological chemistry Vol. 273; no. 4; pp. 2312 - 2321
Main Authors:	Osterwalder, Thomas, Cinelli, Paolo, Baici, Antonio, Pennella, Amedea, Krueger, Stefan Robert, Schrimpf, Sabine Petra, Meins, Marita, Sonderegger, Peter
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 23-01-1998 American Society for Biochemistry and Molecular Biology
Subjects:	Amino Acid Sequence Animals Axons - metabolism Chick Embryo Fibrinolysin - antagonists & inhibitors Genetic Vectors Glycoproteins - metabolism Immunoglobulin kappa-Chains - genetics Kinetics Mice Molecular Sequence Data Neuropeptides - metabolism Neuroserpin Plasminogen Activators - antagonists & inhibitors Protein Conformation Recombinant Proteins - metabolism Sequence Alignment Serine Proteinase Inhibitors - metabolism Serpins - metabolism Thrombin - antagonists & inhibitors Tissue Plasminogen Activator - antagonists & inhibitors Tumor Cells, Cultured Urokinase-Type Plasminogen Activator - antagonists & inhibitors
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Summary:	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-κ-variable region, an Ig-κ enhancer, and the exon encoding the Ig-κ constant region (Cκ) and produced recombinant neuroserpin as a wild-type protein or as a fusion protein with Cκ. 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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0021-9258 1083-351X
DOI:	10.1074/jbc.273.4.2312