Purification and characterization of a trophic factor for embryonic peripheral neurons: comparison with fibroblast growth factors.
The validation of NGF as a physiologically important neurotrophic factor has led to intense efforts to identify novel polypeptide growth factors for neurons. We report here the details of a greater than 80,000-fold purification of a neurotrophic molecule, referred to as growth-promoting activity (GPA), from chicken sciatic nerves. The final product of the purification migrated as a protein band of 21.5 kd, its apparent pI was approximately 4.8, and the ED50 of the most active preparation was approximately 10 pg/ml. Amino acid sequence of a proteolytic digestion fragment of GPA revealed homology with the recently published sequences for rabbit and rat sciatic nerve CNTF. Thus this molecule may be the chicken form of CNTF. Analysis of the specificity of action of GPA showed that, in addition to E8 ciliary ganglion neurons, the factor was able to support short-term survival of E8 dorsal root ganglion and E12 sympathetic neurons. This range of specificities of biological action was also seen with both acidic and basic FGF in the presence of heparin. The biological activity of GPA differed from that of FGF in that it was not potentiated by heparin and did not stimulate mitogenesis in chick fibroblasts.
Eckenstein
F P
FP
Department of Cell Biology and Anatomy, Oregon Health Sciences University, Portland 97201.
Esch
F
F
Holbert
T
T
Blacher
R W
RW
Nishi
R
R
D000595:Amino Acid Sequence; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002645:Chickens; D005346:Fibroblast Growth Factors / Q000032:analysis; D005724:Ganglia / Q000166:cytology; D006133:Growth Substances / Q000302:isolation & purification* / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009474:Neurons / Q000166:cytology*; D012584:Sciatic Nerve / Q000032:analysis
D006133:Growth Substances; D005346:Fibroblast Growth Factors