title: Conformational changes of brain-derived neurotrophic factor during reversed-phase high-performance liquid chromatography.
abstract: Recombinant human brain-derived neurotrophic factor (r-HuBDNF) is eluted as two peaks under reversed-phase liquid chromatographic conditions with gradient elution. Sodium dodecyl sulfate polyacrylamide gel electrophoresis confirmed identical molecular weights in the two peaks, while rechromatography of the separated peaks showed interconvertibility. The two peaks are identified as the monomeric forms of the parent molecules. The molecular weight of the components in the peaks was determined by on-line 90 degree light scattering using a fluorescence detector as a scatterograph. The early eluted peak is a folded form of the r-HuBDNF monomer, while the later eluted peak is an unfolded form of the BDNF monomer. The conformational states were established using a fluorescence detector both at a fixed wavelength and in the scanning mode.
authors.affiliation: AMGEN, Inc., Amgen Center, Thousand Oaks, CA 91320-1789.
authors.forename: R
authors.initials: R
authors.lastname: Rosenfeld
authors.forename: K
authors.initials: K
authors.lastname: Benedek
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor; D016466:CHO Cells; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D019791:Guanidine; D006146:Guanidines / Q000737:chemistry; D006801:Humans; D008027:Light; D009414:Nerve Growth Factors / Q000737:chemistry*; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D006146:Guanidines; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D019791:Guanidine