Part:BBa_K4165048

HtrA1 Switch 28

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), GSGSG Linker (BBa_K4165066), TD28REV (BBa_K4165006), GGSGGGGG Linker (BBa_K4165019), WWW (BBa_K4165007), GSGSG Linker (BBa_K4165066), WAP inhibitor (BBa_K4165008), and T7 terminator (BBa_K731721).

Usage and Biology

Switch 28 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 switch to see the final structure of the switch, the top model which has score 4 out of 6 from RosettaFold.

                                Figure 1. The 3D structure of switch 28 modeled by RosettaFold. 

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 532
  Illegal AgeI site found at 268
  
• 1000
  COMPATIBLE WITH RFC[1000]