Part:BBa_K4602001
HPV16_L1 (HPV16 major capsid protein)
Coding sequence of HPV16 (human papilloma virus) L1 major capsid protein. For expression in E. coli.
Profile
Name: HPV16_L1
Base Pairs: 1518 bp
Peptide length: 505 AAs
Molar mass : 56277.76 Da
Origin: Human papillomavirus type 16
Original function: major capsid protein L1 [Human papillomavirus type 16]
The HPV L1 used for this project:
His-sfGFP-TEV linker fusion
Base Pairs: 2316 bp
Peptide length: 771 AAs
Molar mass: 86092.08 Da
History on HPV
HPV or human papilloma virus is a DNA virus of the Papillomaviridae family that can infect humans. About 200 different genotypes are known. They can be transmitted through the skin and/or mucous membranes. The most studied genotype is the HPV 16 because it is responsible for most cancert cause by an HPV. it was observed for the first time by Georgios Papanicolaou in 1925 and indentified by Harald zur Hausen at the end of the twentieth century
Literature overview :
The L1 protein is one of the two major proteins of the HPV capsid along with the L2 protein. On the capsid, the L1 protein is structured as a pentree. In addition, this protein has the capacity to spontaneously self-assemble into virus-like particles (VLP) after having been synthesized and purified in vitro.
Usage and Biology
In our project we try to synthesize a protein withe AI that binds to HPV. For this purpose we have purified, by his-TAG column, the protein L1 of HPV 16 wtihe a GFP a His tag and a TEV site that allows us to remove GFP and His Tag
Also We tested this property with migration on acrylamide gel in native conditions
Native PAGE + SDS PAGE :
Figure 1: we observe that it has a lot of heavier band at the level for protein L1 of HPV 16 in the native page, this indicates that the protein L1 assembles well in pentamer and can potentielement form VLP.
Additional information
AlphaFold 2 prediction for the clean protein
Figure 2: Alphafold 2.0 structure prediction of HPV16_L1 on its own. Structure predicted to fold correctly, but with low confidence on the linker regions. "Very low" confidence tail (red) corresponds to N-terminal 6xHis-tag, while the "low" confidence helical tail corresponds to the TEV-Linker region.
AlphaFold 2 for the GFP fusion protein
Figure 3: Alphafold 2.0 structure prediction of HPV16_L1 with sfGFP. Structure predicted to fold correctly, but with low confidence on the linker regions. "Very low" confidence tail (red) corresponds to N-terminal 6xHis-tag, while the "low" confidence helical tail corresponds to the TEV-Linker region. The presence of the tails is not predicted to affect sfGFP folding. we can see a domain beetween an α-helix of HPV16_L1 and an α-helix of sfGFP who can interact.
InterPro scan
Figure 4: domain recognized by Interpro.
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]
Functional Parameters
| None |