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Study of the Structure and Biological Activity of the Amino-Terminus of the α-Toxin from Clostridium welchii Type A.
- Source :
-
Current microbiology [Curr Microbiol] 2019 Oct; Vol. 76 (10), pp. 1175-1185. Date of Electronic Publication: 2019 Jul 08. - Publication Year :
- 2019
-
Abstract
- To explore the biological activity of Clostridium welchii α-toxin (CPA), the Asp56 residue of CPA was mutated to glycine (CPA D56G) by site-directed mutagenesis, and the 250 amino acid amino-terminal phospholipase C (PLC)-containing domain of CPA (PLC1-250) was isolated. The secondary and three-dimensional (3D) structures of CPA D56G and PLC1-250 were predicted, and the results showed that the secondary structures of CPA D56G and PLC1-250 were composed of α-helices and random coils. The 3D structures of CPA D56G and PLC1-250 were similar to the 3D structures of CPA. The circular dichroism (CD) spectrum of CPA D56G differed from the CD spectrum of CPA, but the CD spectrum of PLC1-250 was similar to the CD spectrum of CPA. Biological activity assays showed that CPA D56G lost the PLC activity of CPA and that mice immunized with CPA D56G were protected against a challenge with 1 MLD C. welchii type A strain C57-1. In addition, PLC1-250 contained the PLC activity of CPA. This study laid a solid foundation for future studies on the relationship between the molecular structure and biological function of CPA and its molecular mechanism. Our study also provided CPA D56G as a candidate strain for engineering a CPA subunit vaccine for C. welchii type A.
- Subjects :
- Amino Acid Sequence
Animals
Antigens, Bacterial administration & dosage
Antigens, Bacterial immunology
Bacterial Toxins genetics
Bacterial Toxins immunology
Calcium-Binding Proteins genetics
Calcium-Binding Proteins immunology
Clostridium Infections pathology
Clostridium Infections prevention & control
Clostridium perfringens immunology
Immunization
Mice
Mutation
Protein Conformation
Structure-Activity Relationship
Type C Phospholipases genetics
Type C Phospholipases immunology
Bacterial Toxins chemistry
Bacterial Toxins metabolism
Calcium-Binding Proteins chemistry
Calcium-Binding Proteins metabolism
Clostridium perfringens chemistry
Type C Phospholipases chemistry
Type C Phospholipases metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1432-0991
- Volume :
- 76
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Current microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 31286181
- Full Text :
- https://doi.org/10.1007/s00284-019-01733-5