proBrain Derived Neurotrophic Factor (WThuman)

 

proBDNF (WT-human)     

proBrain-Derived Neurotrophic Factor (WT-human)

recombinant, E. coli

Product#:

B-257

Sizes:

1 µg

5 x 1 µg

5 µg

M.W.:

51.78 kDa.

Bioassay:

proBDNF (WT-human) was bioassayed for neurotrophic activity in Trk B transfected PC12 cells to developed Neurites outgrowth and by its ability to activate MAPK in Trk B transfected HEK 293 cells.

proBDNF (WT-human) mediates neurite outgrowth in TrkB transfected PC12 cells.  PC12 cells were transiently transfected with TrkB/pcDNA3 containing the green fluorescence protein (GFP) as a reporter. One day post transfection, the cells were stimulated with 20ng/ml proBDNF (WT-human) (#B-257) or 10ng/ml human BDNF (#B-250). Development of neurites was visualized after 6 days using bright light microscopy.

proBDNF (WT-human) mediates ERK 1/2 activation MAPK in TrkB transfected HEK293 cells. HEK293 cells stably expressing TrkB receptor were serum-starved for 2h and then challenged for 10 min in the presence or absence of proBDNF (Mut-human) (#B-256), proBDNF (WT-human) (#B-257), proBDNF (Mut-mouse) (#B-243) or human BDNF (#B-250). Cell proteins were resolved by SDS PAGE and the level of ERK1/2 phosphorylation determined using anti-phosphoERK.

 

 

 

BDNF is a neurotrophic factor produced by proteolytic cleavage of its precursor, proBDNF. The biologically relevant form of the protein was thought to be the mature form, BDNF.¹ The actions of BDNF are mediated via the binding to TrkB or p75.^2,3

The precursor form was thought to be important for the correct folding, secretion and trafficking of the mature protein.  A single-nucleotide polymorphism (Val66 to Met) in the pro-domain of the human BDNF gene impairs intracellular trafficking and regulated secretion of BDNF in primary cortical neurons and neurosecretory cells but not in endothelial and vascular cells.⁴ This has been shown to affect memory and lead to abnormal hippocampal function in humans.⁵

The finding that proBDNF and not mature BDNF is the preferred ligand for p75, has ushered in a new era which reexamines the biological roles of the two forms.⁶ Some biological roles for proBDNF have been proposed. It has been shown to be a pro-apoptotic ligand for sympathetic neurons⁷ expressing both p75 and sortlin, and to be involved in LTD⁸. On the other hand it has also been shown to elicit prototypical TrkB responses in biological assays, such as TrkB tyrosine phosphorylation, and activation of ERK1/2.⁹

In brain homogenates a mixture of both, proBDNF and mature BDNF has been found^10,11  and in cortical neurons secretion of proBDNF has been shown.⁷ Binding of both proBDNF and mature BDNF to TrkB has been proposed to be via the R103 residue in the mature portion.⁹

 

 

Alomone Labs is pleased to offer both wild type and mutated human proBDNF which will serve as important tools in the elucidation of the biological effects of these intriguing proteins.

 

 

 

 

The amino acid sequence for proBDNF (WT-human) (#B-257) is as follows.

 

MAPMKEANIRQGGLAYPGVRTHGTLESNGPKAGSRGLTSLADTFEHVIEENN

KDADLYTSRVMLSSQVPLEPPLLFLLEEYKNYLDAANMSMRVRRHSDPARR GELSVCDSIS EWVTAADKKT AVDMSGGTVT VLEKVPVSKG QLKQYFYETK CNPMGYTKEG CRGIDKRHWN SQCRTTQSYV RALTMDSKKR IGWRFIRIDT SCVCTLTIKR GR