Interactions between carboxypeptidase M and kinin B1 receptor in endothelial cells
Autor: | Rafael Filippelli da Silva, Paola Bianchi Guimarães, Clovis R. Nakaie, João Bosco Pesquero, Jair R. Chagas, Liliam Fernandes, Adriana K. Carmona, Michael Bader, Carolina Caldas Hoff |
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Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Endothelium Immunology Bradykinin GPI-Linked Proteins Receptor Bradykinin B1 Rats Sprague-Dawley 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine parasitic diseases medicine Animals Humans Receptor Lung Cells Cultured Mice Knockout Pharmacology Chemistry Kallidin HEK 293 cells Endothelial Cells Metalloendopeptidases Transfection Kinin Cell biology Mice Inbred C57BL 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis embryonic structures Knockout mouse Rats Transgenic human activities |
Zdroj: | Inflammation Research. 68:845-855 |
ISSN: | 1420-908X 1023-3830 |
Popis: | Carboxypeptidase M (CPM) is a glycosylphosphatidylinositol anchored enzyme that plays an important role in the kallikrein–kinin system (KKS). CPM catalytic domain hydrolyzes Arg from C-terminal peptides (i.e., bradykinin and kallidin), generating des-Arg-kinins, the agonists of B1 receptor (B1R). It is known that CPM and kinin B1R are co-localized in the plasma membrane microdomains, where they interact with each other, facilitating receptor signaling. We hypothesized here that this CPM-B1R interaction could also affect the activity of the enzyme. Thus, in this work, we evaluated the impact of B1R presence or absence on CPM activity and expression, using primary culture of microvascular endothelial cells from wild-type, kinin B1R knockout mice (B 1 −/− ), and transgenic rats overexpressing B1 receptor exclusively in the endothelium. In addition, HEK293T cells, as wells as B 1 −/− primary culture of endothelial cells, both transfected with B1R, were also used. CPM expression and activity were downregulated in cells of knockout mice compared to control and this reduction was rescued after B1R transfection. Cells overexpressing B1R presented higher levels of CPM mRNA, protein, and activity. This profile was reverted by pre-incubation with the B1R antagonist, R715, in highly expressing receptor cells. Our data show that kinin B1R positively modulates both CPM expression and activity, suggesting that CPM-B1R interaction in membrane microdomains might affect enzyme activity, beyond interfering in receptors signaling. This work highlights the interactions among different components of KKS and contributes to a better understanding of its patho-physiological role. |
Databáze: | OpenAIRE |
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