TETANO
TETANO
TETANO
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<strong>TETANO</strong>
<strong>TETANO</strong> NEONATALE
BOTULISMO
TETANUS<br />
SNC<br />
spastic paralysis<br />
BOTULISM<br />
SNP<br />
flaccid paralisys
NMJ STRUCTURE
MECCANISMO DI INTOSSICAZIONE INTRACELLULARE
SNARE COMPLEX
sinapsi di controllo<br />
sinapsi intossicata
IDENTIFICATION OF KEY RESIDUES OF THE ENDOPEPTIDASE<br />
ACTIVITY OF TeNT AND BoNT/A LIGHT CHAINS
SITE-DIRECTED MUTAGENESIS<br />
Ala<br />
Glu 261<br />
in BoNT/A<br />
Glu 271<br />
in TeNT<br />
Asp<br />
Ala<br />
Ala<br />
Phe 265<br />
in BoNT/A<br />
Tyr 365<br />
in BoNT/A<br />
Tyr 375<br />
in TeNT<br />
Ala<br />
Phe<br />
Ala
PROTEOLYTIC ACTIVITY OF TeNT<br />
MUTANTS<br />
% of proteolysis<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
0 1 2 3<br />
time (hours)<br />
LC<br />
Tyr375Phe<br />
Tyr375Ala<br />
% of proteolysis<br />
100<br />
80<br />
60<br />
40<br />
20<br />
0<br />
0 1 2 3<br />
time (hours)<br />
LC<br />
Glu271Asp<br />
Glu271Ala
PROTEOLYTIC ACTIVITY OF BoNT/A<br />
MUTANTS<br />
% of proteolysis<br />
100<br />
90<br />
80<br />
70<br />
60<br />
50<br />
40<br />
30<br />
20<br />
10<br />
0<br />
0 20 40 60 80 100 120<br />
Time (min)<br />
Lc<br />
Glu261Ala<br />
Tyr365Ala<br />
Phe265Ala
CONCLUSIONS OF THE MUTAGENESIS STUDY<br />
- Glu261 in BoNT/A and the correspondant residue in TeNT<br />
is the fourth ligand of the catalytic zinc<br />
- Tyr365 in BoNT/A and the correspondant residue in TeNT<br />
has an important role in assisting the hydrolysis<br />
-Phe266<br />
might have a role in substrate binding