Abstract: The present invention relates to the pharmaceutical field. Specifically, it contemplates a polynucleotide encoding a neurotoxin polypeptide exhibiting a reduced duration of the biological effect in a subject, wherein said polypeptide comprises at least one E3 ligase recognition motif in the light chain, wherein said E3 ligase recognition motif is preferably a binding motif for the E3 ligase MDM2. The invention further pertains to polypeptides encoded by the polynucleotide of the invention as well as polypeptides comprising one or more amino acid substitutions. Further encompassed by the present invention are vectors and host cells comprising the said polynucleotide, polypeptides encoded thereby and antibodies specifically binding to the polypeptides. Moreover, the invention relates to medicaments comprising said polynucleotides and polypeptides, as well as specific therapeutic applications thereof. Furthermore, the present invention contemplates methods for the manufacture of the polypeptides and medicaments.

Abstract: This invention relates to novel recombinant clostridial neurotoxins exhibiting increased membrane localization and to methods for the manufacture of such recombinant clostridial neurotoxins. These methods comprise the steps of inserting a nucleic acid sequence coding for a C2 domain into a nucleic acid sequence coding for a parental clostridial neurotoxin and expression of the recombinant nucleic acid sequence comprising the C2 domain in a host cell. The invention further relates to novel recombinant single-chain precursor clostridial neurotoxins used in such methods, nucleic acid sequences encoding such recombinant single-chain precursor clostridial neurotoxins, and pharmaceutical compositions comprising the recombinant clostridial neurotoxin with increased membrane localization.

Abstract: This invention relates to a novel method for manufacturing and obtaining recombinant proteins, such as clostridial neurotoxins, harbouring an N-terminal lysine from precursor proteins. The method comprises the step of expressing a nucleic acid sequence encoding a precursor protein comprising an N-terminal motif, which can be recognised by an endoprotease specific for a lysine in P?1 position, and the step of cleaving the precursor protein with the endoprotease. The invention further relates to novel precursor proteins used in such methods, nucleic acid sequences encoding such precursor proteins and novel recombinant proteins, such as clostridial neurotoxins, harbouring an N-terminal lysine.

Abstract: The present invention relates to the pharmaceutical field. Specifically, it contemplates a polynucleotide encoding a neurotoxin polypeptide exhibiting a reduced duration of the biological effect in a subject, wherein said polypeptide comprises at least one E3 ligase recognition motif in the light chain, wherein said E3 ligase recognition motif is preferably a binding motif for the E3 ligase MDM2. The invention further pertains to polypeptides encoded by the polynucleotide of the invention as well as polypeptides comprising one or more amino acid substitutions. Further encompassed by the present invention are vectors and host cells comprising the said polynucleotide, polypeptides encoded thereby and antibodies specifically binding to the polypeptides. Moreover, the invention relates to medicaments comprising said polynucleotides and polypeptides, as well as specific therapeutic applications thereof. Furthermore, the present invention contemplates methods for the manufacture of the polypeptides and medicaments.