Structure–function discrepancy in Clostridium botulinum C3 toxin for its rational prioritization as a subunit vaccine

Abstract

Clostridium botulinum is anaerobic pathogenic bacterium causing food-born botulism in human and animals by producing botulinum neurotoxins A-H, C2, and C3 cytotoxins. Physiological group III strains (type C and D) of this bacterium are capable of producing C2 and C3 toxins in cattle and avian. Herein, we have revealed the structure–function disparity of C3 toxins from two different C. botulinum type C phage (CboC) and type D phage (CboD) to design avirulent toxins rationally. Structure–function discrepancy of the both toxins was computationally evaluated from their homology models based on the conservation in sequence–structure–function relationships upon covariation and point mutations. It has shown that 8 avirulent mutants were generated from CboC of 34 mutants while 27 avirulent mutants resulted from CboD mutants. No major changes were found in tertiary structure of these toxins; however, some structural variations appeared in the coiled and loop regions. Correlated mutation on the first residue would disorder or revolutionize the hydrogen bonding pattern of the coevolved pairs. It suggested that the residues coupling in the local structural environments were compensated with coevolved pairs so as to preserve a pseudocatalytic function in the avirulent mutants. Avirulent mutants of C3 toxins have shown a stable structure with a common blue print of folding process and also attained a near-native backrub ensemble. Thus, we concluded that selecting the site-directed mutagenesis sites are very important criteria for designing avirulent toxins, in development of rational subunit vaccines, to cattle and avian, but the vaccine specificity can be determined by the C3 toxins of C. botulinum harboring phages.

Keywords:

• ADP ribosyltransferase
• binary toxins
• vaccine
• molecular evolution
• structure–function integrity
• C3 exoenzyme
• molecular pathogenesis
• virulence

Acknowledgements

Life Science Research Board-Defence Research and Development Organisation (Sanction No. DLS/81/48222/LSRB-249/BTB/2012), New Delhi, India, is duly acknowledged for financial support.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by the Life Science Research Board-Defence Research and Development Organisation [Sanction No. DLS/81/48222/LSRB-249/BTB/2012], New Delhi, India.