Part:BBa_J58105:Design

Synthetic periplasmic binding protein that docks a vanillin molecule

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Unknown
• 21
  INCOMPATIBLE WITH RFC[21]

  Unknown
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Open and closed conformation of synthetic vanilline binding protein designed using an automatic computational design procedure.

We have used a structure for the closed rbsB (D-ribose binding protein, periplasmic) which has an x-ray structure with pdb 2DRI (271 residues). The vanilline molecule has a small structure and is not so different from the TNT and other ligands used by the designs of Hellinga.

Vanilline molecule.

We have considered the 2DRI pdb structure and we have removed all side chains corresponding to the aminoacids surrounding the original ribose. We have used a computational protein design approach [2] to search among all possible sequences the ones that stabilise (in the sense of a folding free energy) the given atomic structure. Our combinatorial search couples the side chain placement problem, with combinatorial mutagenesis and with the docking problem. Therefore, it explores the best binding mode. It also tries to find solutions with all possible ligand-protein h-bonds satisfied, which would confer a high specificity for the ligand.

Source

We have followed the work of Prof. Hellinga [1] but using the methodology of [2].

References

[1] L.L. Looger, M.A. Dwyer, J. Smith, H.W. Hellinga. Computational design of receptor and sensor proteins with novel functions. Nature, 423, 185-190 (2003).

[2] A. Jaramillo, L. Wernisch, S. Hery and S.J. Wodak. Folding free energy function selects native-like protein sequences in the core but not on the surface. Proc. Natl. Acad. Sci. 99 (2002), 13554-13559.