View source for 14-3-3

==14-3-3==

https://medlineplus.gov/genetics/gene/ywhae/

Ywhae gene = 14-3-3 epsilon = tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, epsilon polypeptide (Ywhae)

Mus musculus tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, epsilon polypeptide (Ywhae), mRNA
NCBI Reference Sequence: NM_009536.4

==Dr. Yi Zhou's 14-3-3 FKO (Thy1-YFP-DIfopein) Transgenic Mice==

See SFN abstracts (2010) 53.1/M17 and (2011) 790.09/FF16

From Kourtney Graham's Honor's Thesis:

<blockquote>
In order to study the role of 14-3-3 proteins in the nervous system_ in
vivo_, we have successfully created lines of neuronal-specific
transgenic mice that may inhibit 14-3-3 dependent cellular processes.
These transgenic mice express a YFP-fused peptide (R18) which binds to
endogenous 14-3-3, and thus blocking its capability to interact with its
binding partners; and are therefore considered to be a functional
knockout. The transgenes are expressed under the control of the Thy-1
promoter which is neuronal-specific and produces variation of transgene
expression in different brain regions, which is great among founder
lines but minimal among descendants of a single founder (Feng et al.
2000). To minimize strain-dependent genetic variation, we have
backcrossed the founder lines with wild-type C57BL/6 mice for at least 8
generations.
</blockquote>


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)

==R18 peptide ==

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].

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]


R18 Sequence:

 Pro-His-Cys-Val-Pro-Arg-Asp-Leu-Ser-Trp-Leu-Asp-Leu-Glu-Ala- Asn-Met-Cys-Leu-Pro-OH
 
 PHCVPRDLSWLDLEANMCLP

Difopein sequence (with individual R18 monomers in parentheses):

 SADGA (PHCVP RDLSW LDLEA NMCLP) GAAGL DSADG A(PHCV PRDLS WLDLE ANMCL P) GAAG LE

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.

==Thy-1 Promoter==

PMID 11086982

==YFP==

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).

== 14-3-3 FKO papers==
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.

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

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

==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]

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]

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]