Preparation and properties of recombinant Clostridium ramosum IgA proteinase. Isolation of Fc-SC and Fab fragments of human secretory IgA
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
33895263
DOI
10.1016/j.pep.2021.105891
PII: S1046-5928(21)00074-7
Knihovny.cz E-resources
- Keywords
- Clostridium ramosum, Enzymatic activity, IgA proteinase, Storage conditions,
- MeSH
- Bacterial Proteins chemistry genetics MeSH
- Firmicutes enzymology genetics MeSH
- Immunoglobulin A, Secretory chemistry MeSH
- Immunoglobulin Fab Fragments * chemistry isolation & purification MeSH
- Immunoglobulin Fc Fragments * chemistry isolation & purification MeSH
- Humans MeSH
- Peptide Hydrolases chemistry genetics MeSH
- Recombinant Proteins chemistry genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Bacterial Proteins MeSH
- Immunoglobulin A, Secretory MeSH
- Immunoglobulin Fab Fragments * MeSH
- Immunoglobulin Fc Fragments * MeSH
- Peptide Hydrolases MeSH
- Recombinant Proteins MeSH
Immunoglobulin A (IgA) proteinase from Clostridium ramosum is the enzyme which cleaves IgA of both subclasses; in contrast, the other bacterial proteinases cleave only IgA1 proteins. Previous reports characterized the activity of proteinase naturally secreted by C. ramosum specific for the normal human serum IgA of IgA1 and IgA2m(1) subclasses and also for secretory IgA (SIgA). Its amino acid sequence was determined, and the recombinant proteinase which cleaved IgA of both subclasses was prepared. Here we report the optimized expression, purification, storage conditions and activity testing against purified human milk SIgA. The recombinant C. ramosum IgA proteinase isolated in the high degree of purity exhibited almost complete cleavage of SIgA of both subclasses. The proteinase remained active upon storage for more than 10 month at -20 °C without substantial loss of enzymatic activity. Purified SIgA fragments are suitable for studies of all antigen-binding and Fc-dependent functions of SIgA involved in the protection against infections with mucosal pathogens.
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