The low density lipoprotein receptor-related protein modulates protease activity in the brain by mediating the cellular internalization of both neuroserpin and neuroserpin-tissue-type plasminogen activator complexes.
Proteases contribute to a variety of processes in the brain; consequently, their activity is carefully regulated by protease inhibitors, such as neuroserpin. This inhibitor is thought to be secreted by axons at synaptic regions where it controls tissue-type plasminogen activator (tPA) activity. Mechanisms regulating neuroserpin are not known, and the current studies were undertaken to define the cellular pathways involved in neuroserpin catabolism. We found that both active neuroserpin and neuroserpin.tPA complexes were internalized by mouse cortical cultures and embryonic fibroblasts in a process mediated by the low density lipoprotein receptor-related protein (LRP). Surprisingly, despite the fact that active neuroserpin is internalized by LRP, this form of the molecule does not directly bind to LRP on its own, indicating the requirement of a cofactor for neuroserpin internalization. Our studies ruled out the possibility that endogenously produced plasminogen activators (i.e. tPA and urokinase-type plasminogen activator) are responsible for the LRP-mediated internalization of active neuroserpin, but could not rule out the possibility that another cell-associated proteases capable of binding active neuroserpin functions in this capacity. In summary, neuroserpin levels appear to be carefully regulated by LRP and an unidentified cofactor, and this pathway may be critical for maintaining the balance between proteases and inhibitors.
Makarova
Alexandra
A
Department of Vascular Biology, Holland Laboratory, American Red Cross, Rockville, Maryland 20855, USA.
Mikhailenko
Irina
I
Bugge
Thomas H
TH
List
Karin
K
Lawrence
Daniel A
DA
Strickland
Dudley K
DK
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002460:Cell Line; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D015151:Immunoblotting; D008024:Ligands; D026503:Low Density Lipoprotein Receptor-Related Protein-1 / Q000502:physiology*; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015843:Serpins / Q000378:metabolism*; D013997:Time Factors; D010959:Tissue Plasminogen Activator / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism; D000097522:Neuroserpin
D010450:Endopeptidases; D008024:Ligands; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D009479:Neuropeptides; D011993:Recombinant Fusion Proteins; D015843:Serpins; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D000097522:Neuroserpin