Difference between revisions of "Part:BBa K3515014:Design"
 
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===Design Notes===
 
===Design Notes===
−
Added a Cysteine for immobilization (L664C) opposite to the active site region. Removed C223A, C316A, C370N, C673L, C96S, and C970S. to ensure only one cysteine would bind to an immobilization linker arm. Amino acid substitution considerations were made using an intensive BLAST search to ensure conservation. All disulfides in Alpha-Klotho were kept to ensure proper structural folding.
+
Added a Cysteine for immobilization (L664C) opposite to the active site region. Removed C223A, C316A, C370N, C673L, C963S, and C970S. to ensure only one cysteine would bind to an immobilization linker arm. Amino acid substitution considerations were made using an intensive BLAST search to ensure conservation. All disulfides in Alpha-Klotho were kept to ensure proper structural folding.
  
  

Latest revision as of 18:12, 29 June 2020


Alpha-Klotho Binding Protein with cysteine modification(s) to bind to a biosensor.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1626
    Illegal XhoI site found at 1165
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 160
    Illegal AgeI site found at 987
    Illegal AgeI site found at 1856
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Added a Cysteine for immobilization (L664C) opposite to the active site region. Removed C223A, C316A, C370N, C673L, C963S, and C970S. to ensure only one cysteine would bind to an immobilization linker arm. Amino acid substitution considerations were made using an intensive BLAST search to ensure conservation. All disulfides in Alpha-Klotho were kept to ensure proper structural folding.


Source

The source of this part is its sequence retrieved from the European Nucleotide Archive (M37722.1) along with our own modifications.

References

Chen, G., Liu, Y., Goetz, R., Fu, L., Jayaraman, S., Hu, M.C., Moe, O.W., Liang, G., Li, X. and Mohammadi, M., 2018. α-Klotho is a non-enzymatic molecular scaffold for FGF23 hormone signalling. Nature, 553(7689), pp.461-466.