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Recombinant Endopeptidases IdeS and IdeZ and Their Potential Application
Irina S Boksha1,2, Vladimir G Lunin3, Tatyana A Danilova3
1Gamaleya National Research Center for Epidemiology and Microbiology, Ministry of Health of the Russian Federation, Moscow, 123098, Russia. boksha_irina@mail.ru.
Biochemistry. Biokhimiia
|September 25, 2023
Summary
Streptococcal endopeptidases IdeS and IdeZ, which cleave IgG, were successfully cloned, expressed, and purified. Recombinant IdeZ shows promise for diagnosing equine strangles and other infections.
Area of Science:
- Microbiology
- Biochemistry
- Immunology
Background:
- Endopeptidases IdeS and IdeZ are streptococcal virulence factors that cleave IgG heavy chains.
- These enzymes have potential applications in medicine, veterinary science, and biotechnology.
Purpose of the Study:
- To clone and express genes encoding IdeS and IdeZ in Escherichia coli.
- To purify and characterize the recombinant endopeptidases.
- To evaluate the diagnostic potential of IdeZ in veterinary medicine.
Main Methods:
- Gene cloning and heterologous expression in Escherichia coli.
- Introduction of a 6His-tag for purification.
- Purification using metal affinity chromatography.
- Activity and specificity assays using polyacrylamide gel electrophoresis.
- Application of recombinant IdeZ in immunological analysis of equine infections.
Main Results:
- Successfully cloned and expressed IdeS from Streptococcus pyogenes and IdeZ from Streptococcus zooepidemicus.
- Purified IdeS and IdeZ to apparent homogeneity.
- Demonstrated enzymatic activity against human and animal IgGs.
- Confirmed specificity towards human IgGs.
- Utilized recombinant IdeZ for diagnosing equine strangles and determining antibody titers.
Conclusions:
- Recombinant IdeS and IdeZ are active and specific IgG-degrading endopeptidases.
- IdeZ is a valuable tool for diagnosing infections caused by Streptococcus equi and Streptococcus zooepidemicus.
- The enzymes hold significant potential for applications in medicine, biotechnology, and veterinary diagnostics.
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