The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulation
The complement system plays an important role in host defense and is activated via three different activation pathways. We have previously reported that mannose-binding lectin-associated serine protease (MASP)-3, unlike its splicing variant MASP-1, circulates in an active form and is essential for t...
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| Language: | English |
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Frontiers Media S.A.
2022-08-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.907023/full |
| _version_ | 1818486266092584960 |
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| author | Kohei Kusakari Takeshi Machida Yumi Ishida Tomoko Omori Toshiyuki Suzuki Masayuki Sekimata Ikuo Wada Teizo Fujita Hideharu Sekine |
| author_facet | Kohei Kusakari Takeshi Machida Yumi Ishida Tomoko Omori Toshiyuki Suzuki Masayuki Sekimata Ikuo Wada Teizo Fujita Hideharu Sekine |
| author_sort | Kohei Kusakari |
| collection | DOAJ |
| description | The complement system plays an important role in host defense and is activated via three different activation pathways. We have previously reported that mannose-binding lectin-associated serine protease (MASP)-3, unlike its splicing variant MASP-1, circulates in an active form and is essential for the activation of the alternative pathway (AP) via the activation of complement factor D (FD). On the other hand, like MASP-1 and MASP-2 of the lectin pathway (LP), MASP-3 forms a complex with the pattern recognition molecules (PRMs) of the LP (LP-PRMs). Both MASP-1 and MASP-2 can be activated efficiently when the LP-PRMs complexed with them bind to their ligands. On the other hand, it remains unclear how MASP-3 is activated, or whether complex formation of MASP-3 with LP-PRMs is involved in activation of MASP-3 or its efficiency in the circulation. To address these issues, we generated wild-type (WT) and four mutant recombinant mouse MASP-3 proteins fused with PA (human podoplanin dodecapeptide)-tag (rmMASP-3-PAs), the latter of which have single amino acid substitution for alanine in the CUB1 or CUB2 domain responsible for binding to LP-PRMs. The mutant rmMASP-3-PAs showed significantly reduced in-vivo complex formation with LP-PRMs when compared with WT rmMASP-3-PA. In the in-vivo kinetic analysis of MASP-3 activation, both WT and mutant rmMASP-3-PAs were cleaved into the active forms as early as 30 minutes in the circulation of mice, and no significant difference in the efficiency of MASP-3 cleavage was observed throughout an observation period of 48 hours after intravenous administration. All sera collected 3 hours after administration of each rmMASP-3-PA showed full restoration of the active FD and AP activity in MASP-3-deficient mouse sera at the same levels as WT mouse sera. Unexpectedly, all mutant rmMASP-3-PAs showed faster clearance from the circulation than the WT rmMASP-3-PA. To our knowledge, the current study is the first to show in-vivo kinetics of MASP-3 demonstrating rapid activation and clearance in the circulation. In conclusion, our results demonstrated that the complex formation of MASP-3 with LP-PRMs is not required for in-vivo activation of MASP-3 or its efficiency, but may contribute to the long-term retention of MASP-3 in the circulation. |
| first_indexed | 2024-12-10T16:20:39Z |
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| id | doaj.art-1034958f68c7402a8de6fe5bca250978 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-10T16:20:39Z |
| publishDate | 2022-08-01 |
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| series | Frontiers in Immunology |
| spelling | doaj.art-1034958f68c7402a8de6fe5bca2509782022-12-22T01:41:49ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-08-011310.3389/fimmu.2022.907023907023The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulationKohei Kusakari0Takeshi Machida1Yumi Ishida2Tomoko Omori3Toshiyuki Suzuki4Masayuki Sekimata5Ikuo Wada6Teizo Fujita7Hideharu Sekine8Department of Immunology, Fukushima Medical University, Fukushima, JapanDepartment of Immunology, Fukushima Medical University, Fukushima, JapanDepartment of Immunology, Fukushima Medical University, Fukushima, JapanDepartment of Immunology, Fukushima Medical University, Fukushima, JapanRadioisotope Research Center, Fukushima Medical University, Fukushima, JapanRadioisotope Research Center, Fukushima Medical University, Fukushima, JapanDepartment of Cell Science, Institute of Biomedical Sciences, Fukushima Medical University, Fukushima, JapanFukushima Prefectural General Hygiene Institute, Fukushima, JapanDepartment of Immunology, Fukushima Medical University, Fukushima, JapanThe complement system plays an important role in host defense and is activated via three different activation pathways. We have previously reported that mannose-binding lectin-associated serine protease (MASP)-3, unlike its splicing variant MASP-1, circulates in an active form and is essential for the activation of the alternative pathway (AP) via the activation of complement factor D (FD). On the other hand, like MASP-1 and MASP-2 of the lectin pathway (LP), MASP-3 forms a complex with the pattern recognition molecules (PRMs) of the LP (LP-PRMs). Both MASP-1 and MASP-2 can be activated efficiently when the LP-PRMs complexed with them bind to their ligands. On the other hand, it remains unclear how MASP-3 is activated, or whether complex formation of MASP-3 with LP-PRMs is involved in activation of MASP-3 or its efficiency in the circulation. To address these issues, we generated wild-type (WT) and four mutant recombinant mouse MASP-3 proteins fused with PA (human podoplanin dodecapeptide)-tag (rmMASP-3-PAs), the latter of which have single amino acid substitution for alanine in the CUB1 or CUB2 domain responsible for binding to LP-PRMs. The mutant rmMASP-3-PAs showed significantly reduced in-vivo complex formation with LP-PRMs when compared with WT rmMASP-3-PA. In the in-vivo kinetic analysis of MASP-3 activation, both WT and mutant rmMASP-3-PAs were cleaved into the active forms as early as 30 minutes in the circulation of mice, and no significant difference in the efficiency of MASP-3 cleavage was observed throughout an observation period of 48 hours after intravenous administration. All sera collected 3 hours after administration of each rmMASP-3-PA showed full restoration of the active FD and AP activity in MASP-3-deficient mouse sera at the same levels as WT mouse sera. Unexpectedly, all mutant rmMASP-3-PAs showed faster clearance from the circulation than the WT rmMASP-3-PA. To our knowledge, the current study is the first to show in-vivo kinetics of MASP-3 demonstrating rapid activation and clearance in the circulation. In conclusion, our results demonstrated that the complex formation of MASP-3 with LP-PRMs is not required for in-vivo activation of MASP-3 or its efficiency, but may contribute to the long-term retention of MASP-3 in the circulation.https://www.frontiersin.org/articles/10.3389/fimmu.2022.907023/fullcomplementMASP-3alternative pathwaylectin pathwaypattern recognition molecules |
| spellingShingle | Kohei Kusakari Takeshi Machida Yumi Ishida Tomoko Omori Toshiyuki Suzuki Masayuki Sekimata Ikuo Wada Teizo Fujita Hideharu Sekine The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulation Frontiers in Immunology complement MASP-3 alternative pathway lectin pathway pattern recognition molecules |
| title | The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulation |
| title_full | The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulation |
| title_fullStr | The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulation |
| title_full_unstemmed | The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulation |
| title_short | The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulation |
| title_sort | complex formation of masp 3 with pattern recognition molecules of the lectin complement pathway retains masp 3 in the circulation |
| topic | complement MASP-3 alternative pathway lectin pathway pattern recognition molecules |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.907023/full |
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