Difference between revisions of "Part:BBa K3610007"

Revision as of 08:33, 1 October 2020

EFR Ectodomain without signal sequence

This part is the ectodomain of the plant PRR EFR, a cell surface receptor determining the preception of the bacterial Elongation-Factor Tu.

Usage and Biology

Elongation factor-thermo unstable receptor (EFR) from A. thaliana is a plant pattern-recognition receptor (PRR). It is a cell surface receptor and part of the plants firts defence mechanism against potential pathogens. The EFR receptor is also a leucin-rich-repeats (LRR) receptor-like serine/threonine-protein kinase. The protein consists of an extracellular domain with leucin-rich repeats, a ligand binding domain found in many receptors, a single-pass transmembrane domain and finally an intracellular kinase domain. The ligand binding domain from EFR has high specificity to a bacterial pathogen-associated moleculat pattern (PAMP), namely the epitope elf18 of the abundant protein Elongation Factor Tu (EF-Tu), which is catalyzes the binding of aminoacyl-tRNA (aa-tRNA) to the ribosome in most prokaryotes and therefore is evolutionarily highly conserved. This makes the EFR a receptor that can be activated by the presence of a huge variety of bacteria. Upon binding of the ligand to the extracellular domain, the receptor dimerizes with its coreceptor BRI1-associated receptor kinase (BAK1). This interaction triggers the activation of the intracellular kinase domain of EFR and BAK1, initiating a signal cascade leading to an upregulation of immune response mechanisms. When the ectodomains of EFR and another PRR are swapped, the new chimeric receptors are sometimes still functional (depending on which second receptor is chosen). It has further been shown, that for binding of the ligand and dimerization with BAK1 the extracellular domain and the transmembrane domain are needed. The kinase domain is not necessary for initiating this interaction.

For visualizing the interaction of EFR with BAK1, the cytoplasmic domain can be replaced with a split fluorescent protein or another protein that generates a visual output. In our iGEM project, we fused this sequence to a split-mCherry protein and a split-luciferase in another experiment and then coexpressed the part with BAK1 carrying the counterpart to the split protein instead of the intracellular kinase domain in S. cerevisiae.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 82
Illegal NheI site found at 1012
Illegal NheI site found at 1930
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1598
1000
COMPATIBLE WITH RFC[1000]

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 This part is the ectodomain of the plant PRR EFR, a cell surface receptor determining the preception of the bacterial Elongation-Factor Tu.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+Elongation factor-thermo unstable receptor (EFR) from A. thaliana is a plant pattern-recognition receptor (PRR). It is a cell surface receptor and part of the plants firts defence mechanism against potential pathogens. The EFR receptor is also a leucin-rich-repeats (LRR) receptor-like serine/threonine-protein kinase. The protein consists of an extracellular domain with leucin-rich repeats, a ligand binding domain found in many receptors, a single-pass transmembrane domain and finally an intracellular kinase domain. The ligand binding domain from EFR has high specificity to a bacterial pathogen-associated moleculat pattern (PAMP), namely the epitope elf18 of the abundant protein Elongation Factor Tu (EF-Tu), which is catalyzes the binding of aminoacyl-tRNA (aa-tRNA) to the ribosome in most prokaryotes and therefore is evolutionarily highly conserved. This makes the EFR a receptor that can be activated by the presence of a huge variety of bacteria. Upon binding of the ligand to the extracellular domain, the receptor dimerizes with its coreceptor BRI1-associated receptor kinase (BAK1). This interaction triggers the activation of the intracellular kinase domain of EFR and BAK1, initiating a signal cascade leading to an upregulation of immune response mechanisms.
+When the ectodomains of EFR and another PRR are swapped, the new chimeric receptors are sometimes still functional (depending on which second receptor is chosen). It has further been shown, that for binding of the ligand and dimerization with BAK1 the extracellular domain and the transmembrane domain are needed. The kinase domain is not necessary for initiating this interaction.
+For visualizing the interaction of EFR with BAK1, the cytoplasmic domain can be replaced with a split fluorescent protein or another protein that generates a visual output. In our iGEM project, we fused this sequence to a split-mCherry protein and a split-luciferase in another experiment and then coexpressed the part with BAK1 carrying the counterpart to the split protein instead of the intracellular kinase domain in S. cerevisiae.
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