BBa_K608407
1
BBa_K608407
Precipitator fused with GST tag
2011-09-13T11:00:00Z
2015-05-08T01:12:53Z
The part is constructed from the following protein sequences
(PDB: 3FXI) TLR-4
(PDB: 2z66) N-Terminal Hagfish
(PDB: 2z62) C-Terminal Hagfish
(PDB: 3cvr) bacterial ligase
http://en.wikipedia.org/wiki/Glutathione_S-transferase#GST-tags_and_the_GST_pull-down_assay
Protein domain of Precipitator N-terminally fused to a GST tag domain.
The GST-tag has the size of 220 amino acids (roughly 26 KDa). It is fused to the N-terminus of the precipitator protein BBa_K608406 and includes a PreScission Protease domain for cleavage of the GST tag during protein purification.
http://www.gelifesciences.com/aptrix/upp01077.nsf/content/Products?OpenDocument&parentid=976040&moduleid=38868&zone=Proteomics
Artificial Leucine Rich Repeat(LRR) with C and N-terminal hagfish domain fragments capping the artifical middle part. This part is one version of three different, designed to bind nickel by histidines which are grouped together pointing away from the horseshoe shaped protein.
Bacterial LRR Consensus of the central LRR fragment:
LxxLxLxxNxLxxLPxxLPxx
Protein code:
CPSRCSCSGTEIRCNSKGLTSVPTGIPSS
ATRLELESNKLQSLPHGVFDK
LTQLTKSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LEVLDVSNNHLHSLPDNLPAS
LKELALDTNQLKSVPDGIFDR
LTSLQKIWLHTNPWDCSCPRIDY
LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP
This protein can be used to complex Nickel or Cobalt. The principal mechanism is comparable to Ni-NTA columns, as chelates the ions. Free binding sites of the ions are then exposed, so that a His-tagged protein can attach to them. The design of the protein is of a particular interest, too. LRR are highly conserved motifs throughout evolution. They appear in all kingdoms of life in almost every thinkable role (Ligases, Receptors, Toxins etc.). Their core is highly conserved and provides a very stable backbone, while the intermediate, non-conserved aminoacids are almost freely interchangeable.
This protein can be used to complex up to 4 Nickel or Cobalt. However the principal design oft he protein is of a particular interest, too. LRR are highly conserved motifs throughout evolution. They appear in all kingdoms of life in almost every thinkable role (Ligases, Receptors, Toxins etc.). Their core is highly conserved and provides a very stable backbone, while the intermediate, non-conserved aminoacids are almost freely interchangeable.
We only submitted one of the three versions, to reduce redundancy in the registry. Please contact us for any questions.
false
false
_780_
0
4456
9
In stock
false
Here we investigated an optimal set of non-conserved aminoacids by analysing large sets of similar proteins and databases. You can use this piece of work as a template to design your own protein and give it any function you like, by simply interchanging aminoacids and fusing other domains on the N or C termini.
To guarantee proper folding and to shield off the hydrophobic core, a well studied fragment of an LRR protein coming from hagfish was used. This technique was investigated before
false
R??diger Trojok
annotation2128442
1
C-Terminal Hagfish PDB:2z62
range2128442
1
1371
1447
annotation2128439
1
PreScission Protease cleavage site
range2128439
1
661
687
annotation2128441
1
N-Terminal Hagfish PDB:2z66
range2128441
1
688
745
annotation2128440
1
Precipitator
range2128440
1
688
1447
annotation2128438
1
GST
range2128438
1
1
660
BBa_K608407_sequence
1
atgtcccctatactaggttattggaaaattaagggccttgtgcaacccactcgacttcttttggaatatcttgaagaaaaatatgaagagcatttgtatgagcgcgatgaaggtgataaatggcgaaacaaaaagtttgaattgggtttggagtttcccaatcttccttattatattgatggtgatgttaaattaacacagtctatggccatcatacgttatatagctgacaagcacaacatgttgggtggttgtccaaaagagcgtgcagagatttcaatgcttgaaggagcggttttggatattagatacggtgtttcgagaattgcatatagtaaagactttgaaactctcaaagttgattttcttagcaagctacctgaaatgctgaaaatgttcgaagatcgtttatgtcataaaacatatttaaatggtgatcatgtaacccatcctgacttcatgttgtatgacgctcttgatgttgttttatacatggacccaatgtgcctggatgcgttcccaaaattagtttgttttaaaaaacgtattgaagctatcccacaaattgataagtacttgaaatccagcaagtatatagcatggcctttgcagggctggcaagccacgtttggtggtggcgaccatcctccaaaatcggatctggaagttctgttccaggggcccatgtgtccttctcgttgtagctgttccggaactgaaattcgctgcaattcaaaaggtttgacctcagtcccgacgggtattccgtcaagcgcgacccgtctcgagcttgagagcaataaattgcagagcctccccgacggcgtattcgataagctgacccagctcacgaaatcaaacaaccacttgcattcgctgccggacaatctgccagcgtctcttgaagtattagatgtgtcgaacaaccacctccacagtttgccggacaacttaccggctagcttagaagttctggatgtctctaataaccatctgcactccctgccagacaacttgcctgcatcactggaagtgctcgatgtgagtaacaatcatttacacagtttacctgataacctgcctgccagccttgaagtgcttgacgtaagcaataaccatttacattcgttacctgataatttgcccgcgagcttagaagtgttagacgttagtaacaatcacctgcatagcctgccagataatcttccggcaagtttaaaggaattagcccttgatacgaatcatctgcattcggttcccgacgggatctttgatcgcctgacatcgttacagaaaatttggcttcagactaacccatgggattgctcctgcccccgtatcgattatctcagtcgttggttgaataaaaattctcaaaaggaacaaggttcagccaaatgttcgggctctggcaaaccggtccgctccatcatttgcccgtaataat
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
Chris J. Myers
James Alastair McLaughlin
2017-03-06T15:00:00.000Z