title: Recombinant GDNF: tetanus toxin fragment C fusion protein produced from insect cells.
abstract: Glial cell line-derived neurotrophic factor (GDNF) has potent survival-promoting effects on CNS motor neurons in experimental animals. Its therapeutic efficacy in humans, however, may have been limited by poor bioavailability to the brain and spinal cord. With a view toward improving delivery of GDNF to CNS motor neurons in vivo, we generated a recombinant fusion protein comprised of rat GDNF linked to the non-toxic, neuron-binding fragment of tetanus toxin. Recombinant GDNF:TTC produced from insect cells was a soluble homodimer like wild-type GDNF and was bi-functional with respect to GDNF and TTC activity. Like recombinant rat GDNF, the fusion protein increased levels of immunoreactive phosphoAkt in treated NB41A3-hGFRalpha-1 neuroblastoma cells. Like TTC, GDNF:TTC bound to immobilized ganglioside GT1b in vitro with high affinity and selectivity. These results support further testing of recombinant GDNF:TTC as a non-viral vector to improve delivery of GDNF to brain and spinal cord in vivo.
authors.affiliation: Cecil B. Day Laboratory for Neuromuscular Research, Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors.forename: Jianhong
authors.initials: J
authors.lastname: Li
authors.forename: Ru-Ju
authors.initials: RJ
authors.lastname: Chian
authors.forename: Ilknur
authors.initials: I
authors.lastname: Ay
authors.forename: Samuel A
authors.initials: SA
authors.lastname: Celia
authors.forename: Brenda B
authors.initials: BB
authors.lastname: Kashi
authors.forename: Eric
authors.initials: E
authors.lastname: Tamrazian
authors.forename: Jonathan C
authors.initials: JC
authors.lastname: Matthews
authors.forename: Mary P
authors.initials: MP
authors.lastname: Remington
authors.forename: R Blake
authors.initials: RB
authors.lastname: Pepinsky
authors.forename: Paul S
authors.initials: PS
authors.lastname: Fishman
authors.forename: Robert H
authors.initials: RH
authors.lastname: Brown
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Francis
mesh_terms: D000818:Animals; D002460:Cell Line; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000096:biosynthesis* / Q000235:genetics; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000235:genetics; D014176:Protein Biosynthesis; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism; D013744:Tetanus Toxin / Q000096:biosynthesis* / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D010446:Peptide Fragments; D011994:Recombinant Proteins; D013744:Tetanus Toxin; C040635:tetanus toxin fragment C