Part:BBa_K257018:Design
AIDA-I autotransporter, linker + translocator domains
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1118
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
AIDA-I is the adhesin involved in diffuse adherence of E.coli. It belongs to the monomeric autotransporter protein family of the type-V secretion pathway. AIDA-I is synthesized from the aidA gene as a 145 kDa pre-proprotein with three domains : first, an N-terminal sec-dependent signal sequence that allows transport across the inner membrane; second, a cenral domain of 100kDa, the mature AIDA-I, bearing the functional part of the protein; and finally, a C-terminal membrane-embedded domain of 45kDa, AIDAc, which is predicted to form a pore in the outer membrane and might serve as a translocation conduit for mature AIDA-I.
We had to deal with considerations about the linker domain which links the translocated protein to the translocator domain of AIDA-I. In fact, this linker needs to have a minimal length so that the protein translocated would be really functional at the outer side of the outer membrane.
AIDA Translocator domain has been designed using a fusion silver standard 23 prefix in order to be able to make a fusion protein with the N terminal extremity of AIDA translocator. The protein passenger should have both standard 23 (silver fusion) suffix and prefix to allow us to build the fusion protein as follows : sequence signal + passenger + aida translocator
Source
AIDA has been synthesized in vitro.