Nerve growth factor from Vipera lebetina venom.
Nerve growth factor was isolated from the Vipera lebetina venom by a four-step procedure including gel filtration, ion exchange, heparin and hydrophobic chromatography. The purified protein is a glycosylated non-covalently bound homodimer with monomeric molecular mass of 14,380 Da. The cDNA encoding NGF is cloned and sequenced. The amino acid sequence translated from the cDNA comprises 117 or 119 amino acids depending on the N-terminus (truncated or not). The recombinant NGF (expressed in Escherichia coli) was used to prepare the anti-NGF antiserum. The antiserum interacted with the wild-type NGF and enabled to localize NGF during the purification procedure in parallel with MALDI-TOF analysis of tryptic peptides. The isolated NGF caused neurite outgrowth from PC12 cells in concentrations beginning from 2.5 ng/ml.
Paalme
Viiu
V
Tallinn University of Technology, Institute of Gene Technology, 12618 Tallinn, Estonia.
Trummal
Katrin
K
Samel
Mari
M
Tõnismägi
Külli
K
Järvekülg
Lilian
L
Vija
Heiki
H
Subbi
Juhan
J
Siigur
Jüri
J
Siigur
Ene
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002850:Chromatography, Gel; D017931:DNA Primers; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000633:toxicity; D016716:PC12 Cells; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000633:toxicity; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014757:Viper Venoms / Q000737:chemistry*
D017931:DNA Primers; D018076:DNA, Complementary; D009414:Nerve Growth Factors; D011994:Recombinant Proteins; D014757:Viper Venoms