Part:BBa_K4897010

TurboID-FGB

What is it?

TurboID was fused with human beta-fibrinogen which can target the P. acne's surface protein. The interaction of human beta-fibrinogen with P. acne surface protein will lead to the proximity of the Biotinylation of P. acne surface proteins. Thus, the biotinylation of surface proteins will block the signaling and quorum sensing. 

P. acne can attach to human skin and cause infection by traveling from the surface to its infection site. When it reach the infection site and bind to human epithelial cells, this process is irreversible and triggers a series of reaction, initiating bacterial colonization, production of reactive oxygen species, and inflammatory reaction. Thus, preventing P. acne from binding human cells is a crucial mission of BS_United China_2023 to suppress P. acne’s virulence. Specifically, P. acne binds to a blood plasma protein called fibrinogen (FBG). The surface of P. acne contains largely a certain protein, in size of 58 kDa which binds strongly to the N-terminal Fg1 peptide, one of the components of beta-fibrinogen. Thus, P. acne strongly binds to human fibrinogen through its surface protein which we call fibrinogen binding protein. 

Modifying our last year’s project, TurboID, we successfully adopted it to target the P. acne surface protein, the fibrinogen binding protein. FGB would direct TurboID to bind a large number of biotin onto the surface proteins of P. acne when FGB binds to the fibrinogen-binding protein of P. acne.

Usage and Biology

Fig. 1. The Components of TurboID-FGB

As FGB directs TurboID toward the fibrinogen binding protein of P. acne, TurboID will catalyze biotin and ATP it carries to produce Biotinyl 5’-AMP for protein labeling. Within a maximum range of 10 nanometers, TurboID is able to label P. acne’s surface proteins with a large amount of Biotinyl5’-AMP which blocks most of the surface protein of P. acne. As a result, P. acne’s communication with the outside world is completely cut off: the metabolic waste cannot be exported out of the cell, and nutrients cannot be transported into the cell. P. acne is suppressed by TurboID-FGB.

Characterization

Fig. 1. TurboID-FGB staining P. acne.

We can notice that TurboID-FGB can successfully label activated biotin onto P. acne, as the red Streptavidin-568 can strongly binds to activated biotin and colors the P. acne which is labeled with abundant biotin.

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