Part:BBa_K2368003:Experience

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Applications of BBa_K2368003

In our experiment, this part is used to accept the signal inputted in our whole pathway.

Part 1: Dry experiments of T1R2

1、Structure models

Purpose

In order to confirm whether this “radar” T1R2/T1R3 can “sense” the sweetness of different sweetener, we simulate the receptor structure model. It helps us to understand how the sweeteners binding to T1R2/T1R3 receptor visually. Otherwise we hope to find some unknown sweeteners binding sites based on this model, even some ideal but unknown sweeteners.

Method

Initially, to make the signal input more accurate and reliable, we simulate the T1R2/T1R3 receptor structure model using SWISS-MODEL. Additionally, we prepare some sweeteners’ PDB file based on Chemdraw 2D and Chemdraw 3D. And the docking process is performed by using Autodock Vina.

Result

We used homology modeling method to get the structure of human sweet receptorT1R2/T1R3. The quality of the simulate structure within normal range and it only contain the ligand-binding-domain based on the crystal protein structure of Mice.

Fig. 2 The simulate structure of human sweetness receptors ligand-binding domain (LBD)

Then we use software Chemdraw 2D and Chemdraw 3D to build PDB files of some classic sweeteners.

Fig. 3 The 3D structure of some sweeteners

The docking process is carried out under Autodock Vina (Fig. 4-6).

Fig. 4 The docking result of different sweeteners

Fig. 5 The docking result of aspartame

Fig. 6 The docking result of stevioside

2、T1R2/T1R3 receptor activation part

The part is about the sweeteners binding to the receptor and the GPCR produce the initial signal through changing structure. For T1R2/T1R3, we divide the process of inducing signal into four conditions of T1R2/T1R3 as show in Fig. 7. And the reaction between different conditions of protein is used ODEs to describe it. The all ODEs are shown follow.

Fig. 7 T1R2/T1R3 receptor activation reaction diagram

(T1R2/3: the T1R2/T1R3 heterodimer)

The parameters of this part are listed in the Table 1.

The output of the T1R2/T1R3 receptor activation part is showed. (Fig.8)

Fig. 8 The output signal, activated state of T1R2/T1R3, induced by different concentration ligand

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