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| ==14-3-3== | | ==14-3-3== |
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− | http://ghr.nlm.nih.gov/gene/YWHAE
| + | https://medlineplus.gov/genetics/gene/ywhae/ |
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| Ywhae gene = 14-3-3 epsilon = tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, epsilon polypeptide (Ywhae) | | Ywhae gene = 14-3-3 epsilon = tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, epsilon polypeptide (Ywhae) |
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| NCBI Reference Sequence: NM_009536.4 | | NCBI Reference Sequence: NM_009536.4 |
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− | | + | ==Dr. Yi Zhou's 14-3-3 FKO (Thy1-YFP-DIfopein) Transgenic Mice== |
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− | ==Dr. Yi Zhou's 14-3-3 FKO (Thy1-YFP-R18) Transgenic Mice== | |
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| See SFN abstracts (2010) 53.1/M17 and (2011) 790.09/FF16 | | See SFN abstracts (2010) 53.1/M17 and (2011) 790.09/FF16 |
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| From Sun et al PMID 23861400 : | | From Sun et al PMID 23861400 : |
− | Glutamatergic kainate receptors (KAR) KAR-EPSCs at mossy fiber-CA3 synapses decay significantly faster in the 14-3-3 functional knock-out mice (but no difference in NMDA-R or AMPA-R EPSC in same synapses) | + | Glutamatergic kainate receptors (KAR) KAR-EPSCs at mossy fiber-CA3 synapses decay significantly faster in the 14-3-3 functional knock-out mice (but no difference in NMDA-R or AMPA-R EPSC in same synapses) |
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| + | ==R18 peptide == |
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− | ==R18 peptide ==
| + | PMID 11577088 |
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− | PMID 10493820 PMID 11577088 | + | R18 is a synthetic 21 amino acid unphosphorylated peptide (MW 2309.6) which binds to the same amphipathic groove on the surface of 14-3-3 as phosphorylated peptides, with high affinity for all 7 mammalian isoforms of 14-3-3 PMID 10493820. Commercial source of peptide [https://www.enzolifesciences.com/BML-P214/r18-peptide/ Enzo #BML-P214-0001]. |
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− | R18 (aka difopein) is a synthetic unphosphorylated peptide (MW 2309.6) which binds to the same amphipathic groove on the surface of 14-3-3 as phosphorylated peptides, with high affinity for all 7 mammalian isoforms of 14-3-3. | + | Difopein is a construct with two R18 monomers in a single 62 amino acid peptide PMID 11577088. Commercial source of peptide: [https://www.tocris.com/products/difopein_2145 Tocris #1245] |
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− | Sequence:
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− | Pro-His-Cys-Val-Pro-Arg-Asp-Leu-Ser-Trp-Leu-Asp-Leu-Glu-Ala- Asn-Met-Cys-Leu-Pro-OH
| + | R18 Sequence: |
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| + | Pro-His-Cys-Val-Pro-Arg-Asp-Leu-Ser-Trp-Leu-Asp-Leu-Glu-Ala- Asn-Met-Cys-Leu-Pro-OH |
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| PHCVPRDLSWLDLEANMCLP | | PHCVPRDLSWLDLEANMCLP |
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− | Commercial source of peptide [Enzo #BML-P214-0001 http://www.enzolifesciences.com/BML-P214/r18-peptide/]
| + | Difopein sequence (with individual R18 monomers in parentheses): |
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| + | SADGA (PHCVP RDLSW LDLEA NMCLP) GAAGL DSADG A(PHCV PRDLS WLDLE ANMCL P) GAAG LE |
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| + | C54 (consisting of the carboyl-terminal 54-amino acid region of theHMG-box binding protein Ste11p ) is another 14-3-3 inhibitor that is structurally unrelated to R18/difopein. The C54 region permits sequestration of Ste11p in the cytoplasm to inhibit nuclear localization PMID 12697825. |
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| ==Thy-1 Promoter== | | ==Thy-1 Promoter== |
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| ==YFP== | | ==YFP== |
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− | YFP is 239 amino acids long (717 bp) according to plasmid map of Invitrogen [http://www.lifetechnologies.com/order/catalog/product/V35820 Vivid ColorsTM pcDNATM/6.2 N-terminal EmGFP/YFP-DEST vectors] see p, 28 of the manual. R18 is 21 amino acids long (63 bp), so total YFP-R18 fusion protein should be ~ 260 amino acids (780 bp). | + | YFP is 239 amino acids long (717 bp) according to plasmid map of Invitrogen [http://www.thermofisher.com/order/catalog/product/V35820 Vivid ColorsTM pcDNATM/6.2 N-terminal EmGFP/YFP-DEST vectors] see p, 28 of the manual. Difopein is 62 amino acids long (186 bp), so total YFP-difopein fusion protein should be ~ 301 amino acids (903 bp). |
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| == 14-3-3 FKO papers== | | == 14-3-3 FKO papers== |
− | Vogt, Peggy, [http://diginole.lib.fsu.edu/uhm/42/ "Exploring the Function of 14-3-3 Proteins in Neurodegenerative Disease"] (2011). Honors Theses. Paper 42. | + | Vogt, Peggy, [https://diginole.lib.fsu.edu/islandora/object/fsu:204445 "Exploring the Function of 14-3-3 Proteins in Neurodegenerative Disease"] (2011). Honors Theses. Paper 42. |
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| C. Sun, H. Qiao, Q. Zhou, Y. Wang, Y. Wu, Y. Zhou, et al., Modulation of GluK2a Subunit-containing Kainate Receptors by 14-3-3 Proteins, J Biol Chem. 288 (2013) 24676–24690. PMID 23861400 | | C. Sun, H. Qiao, Q. Zhou, Y. Wang, Y. Wu, Y. Zhou, et al., Modulation of GluK2a Subunit-containing Kainate Receptors by 14-3-3 Proteins, J Biol Chem. 288 (2013) 24676–24690. PMID 23861400 |
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| Qiao H, Foote M, Graham K, Wu Y, Zhou Y. 14-3-3 proteins are required for hippocampal long-term potentiation and associative learning and memory. J Neurosci. 34(2014):4801-8. PMID 24695700 | | Qiao H, Foote M, Graham K, Wu Y, Zhou Y. 14-3-3 proteins are required for hippocampal long-term potentiation and associative learning and memory. J Neurosci. 34(2014):4801-8. PMID 24695700 |
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| + | ==Antisera for IP Experiments== |
| + | anti-pan-14-3-3 (K-19) , rabbit polyclonal, Santa Cruz #sc-629, 200 ug/1 ml, $279 [https://www.scbt.com/p/pan-14-3-3-antibody-k-19 datasheet] |
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| + | anti-HDAC4 (H-92), rabbit polyclonal, Santa Cruz #sc-11418, 200 ug/1 ml, $279 [https://www.scbt.com/p/hdac4-antibody-h-92 datasheet] |
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| + | anti-Torc1 (C71D11), rabbit mAb, Cell Signaling #2587 $225 [https://www.cellsignal.com/products/primary-antibodies/torc1-crtc1-c71d11-rabbit-mab/2587?Ntk=Products&Ntt=2587 datasheet] |