proBrain Derived Neurotrophic Factor (mut-mouse)
- 貨號 Cat. No.
- B-243
- 規格 Size
- 1 μg 5x1 μg 5 μg
Description
proBrain Derived Neurotrophic Factor (mut-mouse)
(proBDNF mut-m)
| Product#: | B-243 |
| Sizes: |
1 ug |
| 5x1 ug |
5 ug |
| Preparation: |
proBDNF(mut-m) is a recombinant growth factor produced in E.coli. |
| M.W.: |
| 26.0 kDa for the monomer; in solution proBDNF(mut-m) is found as a dimer of 52.0 kDa. |
| Purity: |
HPLC purified; 99% pure as estimated by SDS-PAGE and mass spectral analysis. |
| Bioassay: |
 | proBDNF (mut-m) (#B-243) and proBDNF (WT-m) (#B-240) mediate neurite outgrowth in TrkB transfected PC12 cells. PC12 cells were transiently transfected with TrkB/GFP pcDNA3. One day post transfection, the cells were stimulated with 0.2 nM hBDNF (#B-250), 0.2 nM proBDNF (WT-m) and 0.2 nM proBDNF (mut-m). Development of neurites was visualized by fluorescence microscopy after 4 days. |
 | proBDNF (WT-m) (#B-240) and proBDNF (mut-m) (B#243) induce ERK1/2 MAPK phosphorylation. HEK-293 cells were transiently transfected with TrkB receptor (pcDNA3-TrkB). Three days post transfection, the cells were serum-starved for 2h and then challenged for 10 min with hBDNF (#B-250), proBDNF(WT-m) or proBDNF(mut-m). Cell proteins were resolved by SDS PAGE and the level of ERK1/2 phosphorylation determined using anti-phosphoERK. |
| References: |
1. | Bibel, M and Barde, Y.A. (2000) Genes&Dev. 14, 2929. |
2. | Cahoon-Metzger, S.M. et al., (2001) Dev.Biol. 232, 246. |
3. | Troy, C.M. et al. (2002) J. Biol. Chem. 277, 34295. |
4. | Chen, Z.Y. et al. (2004) J.Neurosci. 24, 4401. |
5. | Egan, M.F. et al.(2003) Cell 112, 257. |
6. | Lee, R. et al., (2001) Science 294, 1945. |
7. | Teng, H.K. et al.(2005) J. Neurosci. 25, 5455. |
8. | Woo, N.H. et al.(2005) Nat. Neurosci. 8, 1069. |
9. | Fayard, B. et al.2005) J. Neurosci.Res. 80, 18. |
10. | Michalski, B et al. (2003) Mol. Brain Res. 111, 148. |
11. | Zhou, X.F. et al. (2004) J. Neurochem. 91, 704. |
12. | Mowla, S.J. et al. (1999) J. Neurosci. 19, 2069. |
13. | Mowla, S. J. et al. (2001) J. Biol. Chem. 276, 12660. |
14. | Matsumoto, T. et al. (2008) Nat Neurosci, 11, 131. |
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, which has been shown to affect the development of motoneurons, modulate synaptic transmission and affect axonal branching, dendrite growth and synapse number.1 These actions are mediated via the binding of BDNF to TrkB. Different actions are mediated by the binding of BDNF to p75 such as neuronal process retraction2 and neuronal apoptosis.3 The precursor form was thought to be important for the correct folding, secretion and trafficing of the mature protein. A single-nucleotide polymorphism (Val66to Met) in the pro-domain of the human BDNF geneimpairs 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 affects memory and lead to abnormal hippocampal function in humans.5 The finding that proBDNF and not mature BDNF is the preferred ligand for p756 has ushered in a new era which reexamines the biological roles of the two forms. Contradictory biological roles for proBDNFhave 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 shownto elicite prototypical TrkB responses in biological assays, such as TrkB tyrosine phosphorylation, and activation of ERK1/2.9 Binding of both proBDNF and mature BDNF to TrkB has been proposed to be via the R103 residue in the mature portion9 In addition, the question of the relative abundance of the precursor vs the mature form has been investigated. In brain homogenates a mixture of proBDNF and mature BDNF has been found.10,11 The secretion of proBDNF from cortical neurons has been shown.7 Most of the work showing secretion of proBDNF was done in cell lines overexpressing full length BDNF constructs.4, 6,12,13 It has been argued14 that this model misrepresents the authentic situation in vivo, since in the transfected systems the secretion of proBDNF results from overloading the limited capacity of the processing machinery. These authors argue that in fact proBDNF is a transient intermediate and is rapidly converted intracellularly to mature BDNF. Alomone Labs is pleased to offer both wild type and mutated proBDNF which will serve as important tools in the elucidation of the biological effects of these intriguing proteins.
The amino acid sequence for proBDNF (mut-m) (#B-243) is as follows, the residues marked in red has been mutated (R112A, R113G) so that the proBDNF is not cleaved to mature BDNF:
MAPMKEVNVH GQGNLAYPGV RTHGTLESVN GPRAGSRGLT TTSLADTFEH VIEELLDEDQ KVRPNEENHK DADLYTSRVM LSSQVPLEPP LLFLLEEYKN YLDAANMSMR VAGHSDPARR GELSVCDSIS EWVTAADKKT AVDMSGGTVT VLEKVPVSKG QLKQYFYETK CNPMGYTKEG CRGIDKRHWN SQCRTTQSYV RALTMDSKKR IGWRFIRIDT SCVCTLTIKR GR
Alomone Labs is pleased to offer both wild type and mutated proBDNF which will serve as important tools in the elucidation of the biological effects of these intriguing proteins.