PMID- 1729439 OWN - NLM STAT- MEDLINE DCOM- 19920211 LR - 20191101 IS - 0270-6474 (Print) IS - 1529-2401 (Electronic) IS - 0270-6474 (Linking) VI - 12 IP - 1 DP - 1992 Jan TI - NGF/BDNF chimeric proteins: analysis of neurotrophin specificity by homolog-scanning mutagenesis. PG - 306-18 AB - Despite their extensive sequence identities at the amino acid level (approximately 55%), NGF and brain-derived neurotrophic factor (BDNF) display distinct neuronal specificity toward neurons of both the PNS and CNS. To explore which region(s) within these neurotrophic factors might determine their differential actions on various subpopulations of peripheral neurons, a systematic series (homolog-scanning mutagenesis) of chimeric NGF/BDNF molecules was prepared using PCR overlap-extension techniques. After expression in COS-7 cells, the chimeric proteins were tested for their biological activities in neurite outgrowth and neuronal survival assays. This approach led to the functional expression of 12 NGF/BDNF chimeras. Surprisingly, despite replacing successive amino acid segments throughout the entire length of NGF with the corresponding parts of BDNF, all chimeras displayed full NGF-like activity in bioassays carried out with PC12 cells, embryonic chick dorsal root ganglion explants, sympathetic ganglion explants, and dissociated cultures of dorsal root ganglion neurons. Most of the chimeras additionally showed BDNF-like activity as defined by neurite outgrowth on chick nodose ganglion explants. However, none of the chimeras supported the survival of dissociated nodose ganglion neurons. Our results suggest that NGF and BDNF must share very similar higher-order protein structures, and we propose that the overall structure or conformation of NGF, in contrast to short amino acid "active-site" segments, may determine its exact neuronal specificity. FAU - Suter, U AU - Suter U AD - Department of Neurobiology, Stanford University School of Medicine, California 94305-5401. FAU - Angst, C AU - Angst C FAU - Tien, C L AU - Tien CL FAU - Drinkwater, C C AU - Drinkwater CC FAU - Lindsay, R M AU - Lindsay RM FAU - Shooter, E M AU - Shooter EM LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - J Neurosci JT - The Journal of neuroscience : the official journal of the Society for Neuroscience JID - 8102140 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Nerve Growth Factors) RN - 0 (Nerve Tissue Proteins) RN - 0 (Recombinant Fusion Proteins) RN - 9007-49-2 (DNA) SB - IM MH - Amino Acid Sequence MH - Animals MH - Base Sequence MH - Biological Assay MH - Brain-Derived Neurotrophic Factor MH - Cell Line MH - Cell Survival MH - Chickens MH - DNA/chemistry/genetics MH - Ganglia, Spinal/ultrastructure MH - Molecular Sequence Data MH - *Mutagenesis MH - Nerve Growth Factors/chemistry/genetics/*pharmacology MH - Nerve Tissue Proteins/chemistry/genetics/*pharmacology MH - Neurites/physiology MH - Neurons/cytology/physiology MH - Polymerase Chain Reaction MH - Recombinant Fusion Proteins/chemistry/genetics/*pharmacology MH - Structure-Activity Relationship MH - Transfection PMC - PMC6575690 EDAT- 1992/01/01 00:00 MHDA- 1992/01/01 00:01 PMCR- 1992/07/01 CRDT- 1992/01/01 00:00 PHST- 1992/01/01 00:00 [pubmed] PHST- 1992/01/01 00:01 [medline] PHST- 1992/01/01 00:00 [entrez] PHST- 1992/07/01 00:00 [pmc-release] AID - 10.1523/JNEUROSCI.12-01-00306.1992 [doi] PST - ppublish SO - J Neurosci. 1992 Jan;12(1):306-18. doi: 10.1523/JNEUROSCI.12-01-00306.1992.