Difference between revisions of "Part:BBa K4990012"

(→‎What is it)
(→‎How it works)
Line 25: Line 25:
  
 
===How it works===
 
===How it works===
−
The HlpA monomer has been used in designing fusion proteins, as it can bind to HSPG on the surface of colorectal cancer cells. Hence, if you wish to target colorectal cancer cells with a protein, you can design a fusion protein attaching HlpA to one end. If you aim to make a cell target colorectal cancer cells, you can custom design a membrane protein containing HlpA for your chassis cell. Through surface display technology, expressing HlpA on the chassis cell surface would allow it to target colorectal cancer cells [4-6].
+
The HlpA monomer has been used in designing fusion proteins, as it can bind to HSPG on the surface of colorectal cancer cells. Hence, if you wish to target colorectal cancer cells with a protein, you can design a fusion protein attaching HlpA to one end. If you aim to make a cell target colorectal cancer cells, you can custom design a membrane protein containing HlpA for your chassis cell. Through surface display technology, expressing HlpA on the chassis cell surface would allow it to target colorectal cancer cells [2-4].
  
 
<html>
 
<html>
Line 31: Line 31:
 
</html>
 
</html>
  
 +
Examination of the DNA-binding region of Hlp revealed that this area is primarily composed of positively charged residues that form a pocket to accommodate DNA binding. The specific residues that comprise the DNA-binding pocket are depicted in Figure below. It should be noted that the side-chain electron density for Arg56, Lys60, Lys71 and Lys73 of subunit A and Arg54, Lys76 and Lys81 of subunit B was partially disordered and could not be traced in the electrondensity maps. Therefore, the side chains were modeled in idealized rotamer positions for the electrostatic surface calculations.[1]
 +
 +
<html>
 +
<img src="https://static.igem.wiki/teams/4990/wiki/registry/hlpa-r.png" width="450" >
 +
</html>
  
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
  
 +
===Reference===
 +
[1]O'Neil P, Lovell S, Mehzabeen N, et al. Crystal structure of histone-like protein from Streptococcus mutans refined to 1.9 Å resolution[J]. Acta Crystallographica Section F: Structural Biology Communications, 2016, 72(4): 257-262.
 +
 +
[2]Ho C L, Tan H Q, Chua K J, et al. Engineered commensal microbes for diet-mediated colorectal-cancer chemoprevention[J]. Nature biomedical engineering, 2018, 2(1): 27-37.
 +
 +
[3]https://2022.igem.wiki/lzu-china/
 +
 +
[4]Tang, H., Zhou, T., Jin, W., Zong, S., Mamtimin, T., Salama, E. S., ... & Li, X. (2023). Tumor-targeting engineered probiotic Escherichia coli Nissle 1917 inhibits colorectal tumorigenesis and modulates gut microbiota homeostasis in mice. Life Sciences, 324, 121709.
  
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 10:03, 12 October 2023


Tumor-Targeting Rod

Usage in short

You can use it to achieve BL-CRC adhesion.

You need to know

Acurate targeting is one important characteristics for many emerging cancer therapies, that is also what ourproject’s ambition. We designed two fusion proteins, they function like fishingrod, so we gave them the name: Fishing Rod Proteins (FRPs). The one that targetFn names Bacteria Tageting Rod(BTR), the other that target CRC cells names Tumor Tageting Rod(TTR). FRPs are displayed on engineered BL membranes, helping BLtarget to the correct place. Of course, their targets are cancer cells and Fn,rather than fish. Each FRP consists of four parts (Figure1): the signal peptide(white),helping FRPs locate on cell membrane; the membrane protein(pink), the base ofFRPs; and the linker(blue), helping FRPs become flexible; and the targeting fragments(yellow), the most pivotal role in FRPs

What is it

It consists of the GL-BP membrane protein, three repeated G4S linker peptides, and HlpA. Essentially, it involves the surface display technique to express HlpA on the BL membrane for adhesion to HSPG, thereby achieving adhesion between BL and CRC.

How it works

The HlpA monomer has been used in designing fusion proteins, as it can bind to HSPG on the surface of colorectal cancer cells. Hence, if you wish to target colorectal cancer cells with a protein, you can design a fusion protein attaching HlpA to one end. If you aim to make a cell target colorectal cancer cells, you can custom design a membrane protein containing HlpA for your chassis cell. Through surface display technology, expressing HlpA on the chassis cell surface would allow it to target colorectal cancer cells [2-4].

Examination of the DNA-binding region of Hlp revealed that this area is primarily composed of positively charged residues that form a pocket to accommodate DNA binding. The specific residues that comprise the DNA-binding pocket are depicted in Figure below. It should be noted that the side-chain electron density for Arg56, Lys60, Lys71 and Lys73 of subunit A and Arg54, Lys76 and Lys81 of subunit B was partially disordered and could not be traced in the electrondensity maps. Therefore, the side chains were modeled in idealized rotamer positions for the electrostatic surface calculations.[1]


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 358
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 358
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 771
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 358
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 358
    Illegal NgoMIV site found at 1050
    Illegal NgoMIV site found at 1590
  • 1000
    COMPATIBLE WITH RFC[1000]