Characterization and crystallization of recombinant human neurotrophin-4.
Neurotrophin-4 (NT-4) is the most recently discovered member of the neurotrophin family. We have expressed, refolded, and purified recombinant human NT-4 from Escherichia coli and compared it with recombinant human NT-4 secreted into the culture medium of baculovirus-infected insect cells. Both preparations were characterized and determined to be indistinguishable according to several biochemical criteria. Recombinant NT-4 from E. coli was crystallized in a form suitable for x-ray analysis, and characterization of these crystals indicated that NT-4 was present as a dimer within the asymmetric unit. NT-4 was active in promoting the survival of rat TrkB receptor-expressing fibroblasts, but was inactive on embryonic chicken sensory neurons, unlike the other members of the neurotrophin family and in contrast to the reported activities of partially purified NT-4.
Fandl
J P
JP
Regeneron Pharmaceuticals, Inc., Tarrytown, New York 10591.
Tobkes
N J
NJ
McDonald
N Q
NQ
Hendrickson
W A
WA
Ryan
T E
TE
Nigam
S
S
Acheson
A
A
Cudny
H
H
Panayotatos
N
N
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D002942:Circular Dichroism; D003460:Crystallization; D006801:Humans; D009036:Moths; D009414:Nerve Growth Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020801:Receptor, Ciliary Neurotrophic Factor; D017978:Receptors, Growth Factor / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014961:X-Ray Diffraction
D009414:Nerve Growth Factors; D020801:Receptor, Ciliary Neurotrophic Factor; D017978:Receptors, Growth Factor; D011994:Recombinant Proteins; C073922:neurotrophin 4