Linked Life Data

15751983

Source:http://linkedlifedata.com/resource/pubmed/id/15751983

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2005-3-8
pubmed:abstractText
Botulinum neurotoxins (BoNTs A-G) are zinc metalloendoproteases that exhibit extraordinary specificities for proteins involved in neurotransmitter release. In view of the extreme toxicities of these molecules, their applications in human medicine, and potential for misuse, it is of considerable importance to elucidate the mechanisms underlying substrate recognition and to develop inhibitors, with the ultimate goal of obtaining anti-botulinum drugs. We synthesized peptides based on vesicle-associated membrane protein (VAMP) to investigate the substrate requirements of BoNT F, which cleaves VAMP between residues Q58 and K59. The minimum substrate was a peptide containing VAMP residues 32-65, which includes only one of the two VAMP structural motifs thought to be required for botulinum substrate recognition. BoNT F exhibited a strict requirement for residues D57 (P(2)), K59 (P(1)'), and L60 (P(2)'), but peptides containing substitutions for R56 (P(3)), Q58 (P(1)), and S61 (P(3)') were cleaved. Therefore, the P(2), P(1)', and P(2)' residues of VAMP are of paramount importance for BoNT F substrate recognition near the scissile bond. K(i) values of uncleavable analogues were similar to K(m) values of the substrate, suggesting that substrate discrimination occurs at the cleavage step, not at the initial binding step. We then synthesized inhibitors of BoNT F that incorporated d-cysteine in place of glutamine 58, exhibited K(i) values of 1-2 nM, and required binding groups on the N-terminal but not the C-terminal side of the zinc ligand. The latter characteristic distinguishes BoNT F from other zinc metalloendoproteases, including BoNTs A and B.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
44
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4067-73
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed-meshheading:15751983-Amino Acid Sequence, pubmed-meshheading:15751983-Amino Acid Substitution, pubmed-meshheading:15751983-Antitoxins, pubmed-meshheading:15751983-Botulinum Toxins, pubmed-meshheading:15751983-Catalysis, pubmed-meshheading:15751983-Cysteine, pubmed-meshheading:15751983-Humans, pubmed-meshheading:15751983-Hydrolysis, pubmed-meshheading:15751983-Kinetics, pubmed-meshheading:15751983-Membrane Proteins, pubmed-meshheading:15751983-Metalloproteases, pubmed-meshheading:15751983-Molecular Sequence Data, pubmed-meshheading:15751983-Nanotechnology, pubmed-meshheading:15751983-Peptide Fragments, pubmed-meshheading:15751983-Protease Inhibitors, pubmed-meshheading:15751983-Protein Binding, pubmed-meshheading:15751983-R-SNARE Proteins, pubmed-meshheading:15751983-Serotyping, pubmed-meshheading:15751983-Substrate Specificity, pubmed-meshheading:15751983-Sulfhydryl Compounds
pubmed:year
2005
pubmed:articleTitle
Botulinum neurotoxin serotype F: identification of substrate recognition requirements and development of inhibitors with low nanomolar affinity.
pubmed:affiliation
Toxinology Division, United States Army Medical Research Institute of Infectious Diseases, 1425 Porter Street, Fort Detrick, Maryland 21702, USA. james.schmidt@det.amedd.army.mil
pubmed:publicationType
Journal Article