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Characterisation of neurotrophin dimers and monomers

Kolbeck, R. and Jungbluth, S. and Barde, Y. A.. (1994) Characterisation of neurotrophin dimers and monomers. European journal of biochemistry, Vol. 225, H. 3. pp. 995-1003.

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Official URL: http://edoc.unibas.ch/dok/A5259548

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Abstract

When recombinant brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) are purified by reverse-phase chromatography, these neurotrophins elute as two distinct peaks. This is also the case when naturally occurring BDNF is purified from brain tissue. As indicated by gelfiltration experiments, the peaks with shorter retention times correspond to neurotrophin dimers, those with longer retention times to monomers. In contrast, a BDNF mutant with a single amino-acid replacement (Arg-1--<Lys) in the basic processing site common to all neurotrophin precursors elutes as a single peak. This peak is shown by gel-filtration chromatography to consist of dimers with a molecular mass almost twice that of wild-type dimers. N-terminal sequencing indicates an extension of 19 amino acids, including a glycosylated asparagine residue. The biological activity of the BDNF mutant ([R-1K]BDNF) is identical with that of wild-type BDNF when tested in a neuron survival assay. Using this assay, the biological activities of guanidine-hydrochloride-denatured neurotrophin monomers were found to be much lower than that of the dimers, and experiments with NT-3 monomers and NIH3T3 cells expressing trkC suggest that such monomers exist in solution in a conformation that prevents efficient interactions with neurotrophin receptors.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Former Organization Units Biozentrum > Pharmacology/Neurobiology (Barde)
UniBasel Contributors:Barde, Yves-Alain
Item Type:Article, refereed
Article Subtype:Research Article
Bibsysno:Link to catalogue
Publisher:Blackwell
ISSN:0014-2956
Note:Publication type according to Uni Basel Research Database: Journal article
Last Modified:22 Mar 2012 14:21
Deposited On:22 Mar 2012 13:22

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