Part:BBa_K2958008
Fast Acting Single Chain Insulin Analog
This single chain proinsulin analog consists of an A Chain and B Chain from Proinsulin (from BBa_K2417006) and a GGYLPGGGDVGR linker (BBa_K295004). The isoelectric point, or PI, of this insulin is 5.50, as determined by serial cloner.
Description:
Proinsulin is normally made of an A Chain, B Chain, and C chain. Insulin is in its active form after the C chain is cleaved by endopeptidases. Specifically, prohormone convertases PC1 and PC2. However, with the addition of the GGYLPGGGDVGR linker, the insulin-producing process no longer requires this step.This Fast Acting Single Chain Insulin Analog was designed to have a PI of 5.50, the same PI as Insulin Lispro. Modifications include the single chain linker (BBa_K2958004) and a lis-pro amino acid switch at the B28 and B29 position on the B-Chain, which is the same modification as Lispro (unpatented as of 2019). We designed this single chain fast acting insulin with the intent to compare its structure and function to native human insulin in hopes of confirming that the fast acting insulin has a quicker reaction rate than the wild type.
Figure 1. (A) Protein model of native human insulin (active form) produced on Swiss Model software. Orange portion is A chain of insulin, and green portion is B chain of insulin. (B) Protein model of fast acting insulin with GGYLPGGGDVGR linker (BBa_K295005) produced on Swiss Model software. Orange portion is A chain of insulin, green portion is B chain, and red portion is the linker.
Figure 2. Protein model of Native human insulin (orange portion = A chain of native insulin, green portion = B chain of insulin) superimposed on protein model of single chain fast acting insulin (pink portion = A chain of fast acting single chain insulin, blue portion= B chain of fast acting single chain insulin, red portion = GGYLPGGGDVGR linker) produced on Swiss Model Software. Based on these superimposed models alone, it appears that the addition of the linker does not disrupt the structure of the A and B chains. However, the side chains of these insulin molecules require more extensive analysis.
Figure 3.Protein model demonstrating interaction energy (DG) between the single chain native insulin molecule and the ectodomain of the insulin receptor (domain outside of cell membrane). The estimated DG is 1.49372 kcal/mol. In comparison to the estimated DM of our single chain insulin (BBa_K2958006), which was 2.04839 kcal/mol, indicating that the mutations in the long lasting insulin analog decreased the interaction energy by 0.55467 kcal/mol. This model was produced with FoldX software, courtesy of iGEM Team_Moscow 2019.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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