Part:BBa_K4165041

HtrA1 Switch 21

This composite part consists of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), pGS-21a RBS (BBa_K4165016), 6x His-tag (BBa_K4165020), H1A (BBa_K4165000), GGGGS linker (BBa_K4165068), TD28REV (BBa_K4165006), GGSGGGG Linker (BBa_K4165018), Seed Peptide (BBa_K4165012), GGGGS Linker (BBa_K4165068), WAP inhibitor (BBa_K4165008), and T7 terminator (BBa_K731721)

Usage and Biology

Switch 21 plays a role in mediating HTRA1's function. For HTRA1 to become active, a conformational modification in the linker is necessary, which displaces the associated inhibitor from the active site. The conformational rearrangement can be mediated through the affinity clamp for tau and beta-amyloid binding. These clamps are used for stabilizing the inhibitor away from the active site. These two domains (inhibitor and affinity clamp connected with linker1 beside with amyloid beta binding peptide connected to tau binding peptide by linker 2). Additionally, (H1A) binding peptide bound to the PDZ domain and connected to the affinity clamp on the other side with linker3.

Dry lab

Modeling

we modeled this part to see the final structure of the switch, the top model which has score 4 out of 6 from TRrosetta.

                                         Figure 1. The 3D structure of switch 21 modeled by TRrosetta.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 499
  Illegal AgeI site found at 235
  
• 1000
  COMPATIBLE WITH RFC[1000]