Title: Structure-based design of peptide inhibitors of botulinum neurotoxin serotype A proteolytic activity

Authors: Matthew L. Ludivico, Leonard A. Smith, S. Ashraf Ahmed

Addresses: Department of Cell Biology and Biochemistry, Integrated Toxicology Division, US Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, MD 21702, USA. ' Medical Countermeasures Technology, US Army Medical Research and Materiel Command, Fort Detrick, MD 21702-5011, USA. ' Department of Cell Biology and Biochemistry, Integrated Toxicology Division, US Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, MD 21702, USA

Abstract: P1|arginine of the substrate SNAP-25 is an absolute requirement for catalysis, and the active site of botulinum neurotoxin serotype A (BoNT/A) is populated by acidic residues. We reasoned that arginine derivatives, and basic peptides might bind to the proteolytic domain, and act as inhibitors. A systematic investigation of amino acids, derivatives and basic peptides provided proofs of our concept. D-arginine, arginine hydroxamate and basic peptides effectively inhibited BoNT/A activity. Optimisation of the peptide inhibitors resulted in a RRGC tetrapeptide displaying 90% inhibition at 20 microM. Our studies provide an inhibitor scaffold for potential development as a BoNT/A drug candidate.

Keywords: botulinum neurotoxins; BoNT; catalysis; basic tetrapeptides; peptide inhibitors; protease inhibitors; tetrapeptide inhibitors; botulinum neurotoxin inhibitors; arginine hydroxamate; proteolytic activity.

DOI: 10.1504/TBJ.2009.031681

The Botulinum Journal, 2009 Vol.1 No.3, pp.297 - 308

Published online: 17 Feb 2010 *

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