Recombinant GDNF: tetanus toxin fragment C fusion protein produced from insect cells.
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.
Li
Jianhong
J
Cecil B. Day Laboratory for Neuromuscular Research, Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, USA.
Chian
Ru-Ju
RJ
Ay
Ilknur
I
Celia
Samuel A
SA
Kashi
Brenda B
BB
Tamrazian
Eric
E
Matthews
Jonathan C
JC
Remington
Mary P
MP
Pepinsky
R Blake
RB
Fishman
Paul S
PS
Brown
Robert H
RH
Francis
Jonathan W
JW
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
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