Analysis of binding sites on complement factor I that are required for its activity
The central complement inhibitor factor I (FI) degrades activated complement factors C4b and C3b in the presence of cofactors such as C4b-binding protein, factor H, complement receptor 1, and membrane cofactor protein. FI is a serine protease composed of two chains. The light chain comprises the serine protease domain, whereas the heavy chain contains several domains; that is, the FI and membrane attack complex domain (FIMAC), CD5, low density lipoprotein receptor 1 (LDLr1) and LDLr2 domains. To understand better how FI acts as a complement inhibitor, we used homology-based models of FI domains to predict potential binding sites. Specific amino acids were then mutated to yield 16 well expressed mutants, which were then purified from media of eukaryotic cells for functional analyses. The Michaelis constant (K(m)) of all FI mutants toward a small substrate was not altered, whereas some mutants showed increased maximum initial velocity (V(max)). All the mutations in the FIMAC domain affected the ability of FI to degrade C4b and C3b irrespective of the cofactor used, whereas only some mutations in the CD5 and LDLr1/2 domains had a similar effect. These same mutants also showed impaired binding to C3met. In conclusion, the FIMAC domain appears to harbor the main binding sites important for the ability of FI to degrade C4b and C3b.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2010 |
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| Erschienen: |
2010 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:285 |
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| Enthalten in: |
The Journal of biological chemistry - 285(2010), 9 vom: 26. Feb., Seite 6235-45 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Nilsson, Sara C [VerfasserIn] |
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| Links: |
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| Themen: |
80295-43-8 |
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| Anmerkungen: |
Date Completed 26.03.2010 Date Revised 20.10.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1074/jbc.M109.097212 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM194076989 |
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| 100 | 1 | |a Nilsson, Sara C |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Analysis of binding sites on complement factor I that are required for its activity |
| 264 | 1 | |c 2010 | |
| 336 | |a Text |b txt |2 rdacontent | ||
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| 500 | |a Date Completed 26.03.2010 | ||
| 500 | |a Date Revised 20.10.2021 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a The central complement inhibitor factor I (FI) degrades activated complement factors C4b and C3b in the presence of cofactors such as C4b-binding protein, factor H, complement receptor 1, and membrane cofactor protein. FI is a serine protease composed of two chains. The light chain comprises the serine protease domain, whereas the heavy chain contains several domains; that is, the FI and membrane attack complex domain (FIMAC), CD5, low density lipoprotein receptor 1 (LDLr1) and LDLr2 domains. To understand better how FI acts as a complement inhibitor, we used homology-based models of FI domains to predict potential binding sites. Specific amino acids were then mutated to yield 16 well expressed mutants, which were then purified from media of eukaryotic cells for functional analyses. The Michaelis constant (K(m)) of all FI mutants toward a small substrate was not altered, whereas some mutants showed increased maximum initial velocity (V(max)). All the mutations in the FIMAC domain affected the ability of FI to degrade C4b and C3b irrespective of the cofactor used, whereas only some mutations in the CD5 and LDLr1/2 domains had a similar effect. These same mutants also showed impaired binding to C3met. In conclusion, the FIMAC domain appears to harbor the main binding sites important for the ability of FI to degrade C4b and C3b | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Complement C3b |2 NLM | |
| 650 | 7 | |a 80295-43-8 |2 NLM | |
| 650 | 7 | |a Complement C4b |2 NLM | |
| 650 | 7 | |a 80295-50-7 |2 NLM | |
| 650 | 7 | |a Complement Factor I |2 NLM | |
| 650 | 7 | |a EC 3.4.21.45 |2 NLM | |
| 700 | 1 | |a Nita, Izabela |e verfasserin |4 aut | |
| 700 | 1 | |a Månsson, Lisa |e verfasserin |4 aut | |
| 700 | 1 | |a Groeneveld, Tom W L |e verfasserin |4 aut | |
| 700 | 1 | |a Trouw, Leendert A |e verfasserin |4 aut | |
| 700 | 1 | |a Villoutreix, Bruno O |e verfasserin |4 aut | |
| 700 | 1 | |a Blom, Anna M |e verfasserin |4 aut | |
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| 856 | 4 | 0 | |u http://dx.doi.org/10.1074/jbc.M109.097212 |3 Volltext |
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