Furin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10.
Bone morphogenetic protein 10 (BMP10) is a member of the TGF-β superfamily and plays a critical role in heart development. In the postnatal heart, BMP10 is restricted to the right atrium. The inactive pro-BMP10 (∼60 kDa) is processed into active BMP10 (∼14 kDa) by an unknown protease. Proteolytic cl...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2011
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| _version_ | 1797084122725744640 |
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| author | Susan-Resiga, D Essalmani, R Hamelin, J Asselin, M Benjannet, S Chamberland, A Day, R Szumska, D Constam, D Bhattacharya, S Prat, A Seidah, N |
| author_facet | Susan-Resiga, D Essalmani, R Hamelin, J Asselin, M Benjannet, S Chamberland, A Day, R Szumska, D Constam, D Bhattacharya, S Prat, A Seidah, N |
| author_sort | Susan-Resiga, D |
| collection | OXFORD |
| description | Bone morphogenetic protein 10 (BMP10) is a member of the TGF-β superfamily and plays a critical role in heart development. In the postnatal heart, BMP10 is restricted to the right atrium. The inactive pro-BMP10 (∼60 kDa) is processed into active BMP10 (∼14 kDa) by an unknown protease. Proteolytic cleavage occurs at the RIRR(316)↓ site (human), suggesting the involvement of proprotein convertase(s) (PCs). In vitro digestion of a 12-mer peptide encompassing the predicted cleavage site with furin, PACE4, PC5/6, and PC7, showed that furin cleaves the best, whereas PC7 is inactive on this peptide. Ex vivo studies in COS-1 cells, a cell line lacking PC5/6, revealed efficient processing of pro-BMP10 by endogenous PCs other than PC5/6. The lack of processing of overexpressed pro-BMP10 in the furin- and PACE4-deficient cell line, CHO-FD11, and in furin-deficient LoVo cells, was restored by stable (CHO-FD11/Fur cells) or transient (LoVo cells) expression of furin. Use of cell-permeable and cell surface inhibitors suggested that endogenous PCs process pro-BMP10 mostly intracellularly, but also at the cell surface. Ex vivo experiments in mouse primary hepatocytes (wild type, PC5/6 knock-out, and furin knock-out) corroborated the above findings that pro-BMP10 is a substrate for endogenous furin. Western blot analyses of heart right atria extracts from wild type and PACE4 knock-out adult mice showed no significant difference in the processing of pro-BMP10, implying no in vivo role of PACE4. Overall, our in vitro, ex vivo, and in vivo data suggest that furin is the major convertase responsible for the generation of BMP10. |
| first_indexed | 2024-03-07T01:51:01Z |
| format | Journal article |
| id | oxford-uuid:9a155dc3-9f6f-4581-8a1f-0317062dae20 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T01:51:01Z |
| publishDate | 2011 |
| record_format | dspace |
| spelling | oxford-uuid:9a155dc3-9f6f-4581-8a1f-0317062dae202022-03-27T00:19:00ZFurin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:9a155dc3-9f6f-4581-8a1f-0317062dae20EnglishSymplectic Elements at Oxford2011Susan-Resiga, DEssalmani, RHamelin, JAsselin, MBenjannet, SChamberland, ADay, RSzumska, DConstam, DBhattacharya, SPrat, ASeidah, NBone morphogenetic protein 10 (BMP10) is a member of the TGF-β superfamily and plays a critical role in heart development. In the postnatal heart, BMP10 is restricted to the right atrium. The inactive pro-BMP10 (∼60 kDa) is processed into active BMP10 (∼14 kDa) by an unknown protease. Proteolytic cleavage occurs at the RIRR(316)↓ site (human), suggesting the involvement of proprotein convertase(s) (PCs). In vitro digestion of a 12-mer peptide encompassing the predicted cleavage site with furin, PACE4, PC5/6, and PC7, showed that furin cleaves the best, whereas PC7 is inactive on this peptide. Ex vivo studies in COS-1 cells, a cell line lacking PC5/6, revealed efficient processing of pro-BMP10 by endogenous PCs other than PC5/6. The lack of processing of overexpressed pro-BMP10 in the furin- and PACE4-deficient cell line, CHO-FD11, and in furin-deficient LoVo cells, was restored by stable (CHO-FD11/Fur cells) or transient (LoVo cells) expression of furin. Use of cell-permeable and cell surface inhibitors suggested that endogenous PCs process pro-BMP10 mostly intracellularly, but also at the cell surface. Ex vivo experiments in mouse primary hepatocytes (wild type, PC5/6 knock-out, and furin knock-out) corroborated the above findings that pro-BMP10 is a substrate for endogenous furin. Western blot analyses of heart right atria extracts from wild type and PACE4 knock-out adult mice showed no significant difference in the processing of pro-BMP10, implying no in vivo role of PACE4. Overall, our in vitro, ex vivo, and in vivo data suggest that furin is the major convertase responsible for the generation of BMP10. |
| spellingShingle | Susan-Resiga, D Essalmani, R Hamelin, J Asselin, M Benjannet, S Chamberland, A Day, R Szumska, D Constam, D Bhattacharya, S Prat, A Seidah, N Furin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10. |
| title | Furin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10. |
| title_full | Furin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10. |
| title_fullStr | Furin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10. |
| title_full_unstemmed | Furin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10. |
| title_short | Furin is the major processing enzyme of the cardiac-specific growth factor bone morphogenetic protein 10. |
| title_sort | furin is the major processing enzyme of the cardiac specific growth factor bone morphogenetic protein 10 |
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