Difference between revisions of "Part:BBa K3064010"
| Line 10: | Line 10: | ||
Due to the organism, therapeutic antibodies targeting disease-associated antigens in extracellular is a key tool for some certain diseases in current researches. Although some diseases is not caused by virons, trim 21 can be used in these diseased by blocking a particular pathogenic process.<Br/> | Due to the organism, therapeutic antibodies targeting disease-associated antigens in extracellular is a key tool for some certain diseases in current researches. Although some diseases is not caused by virons, trim 21 can be used in these diseased by blocking a particular pathogenic process.<Br/> | ||
| + | |||
| + | ===Result=== | ||
| + | To validate the protein degradation efficiency of the ScFv-based ErbB3 Predator system, we | ||
| + | transfected MCF7 cells with plasmids expressing ScFvErbB3-linker-Fc fusion protein and HA- | ||
| + | Trim21. Western Blotting analysis showed that with the high expression of ScFvErbB3-linker- | ||
| + | Fc fusion protein and HA-Trim21, the ErbB3 protein level decreased dramatically in cells | ||
| + | transfected with ErbB3 Predator (approximately 30%, Fig 1A and B). This result demonstrated that the ScFv-based Predator system could successfully degrade the targeted membrane protein. | ||
| + | |||
| + | https://static.igem.org/mediawiki/parts/c/c6/T--NUDT_CHINA--erbB3-composite.jpg | ||
| + | |||
| + | Figure 1. (A and B) Western blotting assay showing the components of ErbB3 Predator and the degradation of ErbB3. | ||
| + | |||
| + | |||
<!-- --> | <!-- --> | ||
Latest revision as of 00:08, 27 October 2020
mmuTrim21
Trim 21 comes from the family of TRIM. ‘MMu’ means that it is from mouse. It is a known type of E3 ubiquitin-protein ligase as an intracellular antibody effector in the antibody-mediated proteolysis pathway.
Usage and Biology
Trim 21 has a particular domain to recognize the Fc domain and combine with antibody marked no-enveloped virions. Using its E3 ubiquitin-protein ligase activity, trim 21 with the antibody bond to virons is degraded by the Ubiquitin dependent protease degradation pathway.
Due to the organism, therapeutic antibodies targeting disease-associated antigens in extracellular is a key tool for some certain diseases in current researches. Although some diseases is not caused by virons, trim 21 can be used in these diseased by blocking a particular pathogenic process.
Result
To validate the protein degradation efficiency of the ScFv-based ErbB3 Predator system, we transfected MCF7 cells with plasmids expressing ScFvErbB3-linker-Fc fusion protein and HA- Trim21. Western Blotting analysis showed that with the high expression of ScFvErbB3-linker- Fc fusion protein and HA-Trim21, the ErbB3 protein level decreased dramatically in cells transfected with ErbB3 Predator (approximately 30%, Fig 1A and B). This result demonstrated that the ScFv-based Predator system could successfully degrade the targeted membrane protein.
Figure 1. (A and B) Western blotting assay showing the components of ErbB3 Predator and the degradation of ErbB3.
Sequence and Features
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 75
Illegal BamHI site found at 778 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 176
- 1000COMPATIBLE WITH RFC[1000]
References
1.TRIM21[G/OL]. https://en.wikipedia.org/wiki/TRIM21 ,2019.
2.Stian Foss, Ruth Watkinson, Inger Sandlie, et al. TRIM21: a cytosolic Fc receptor with broad antibody isotype specificity[J]. Immunological Reviews, 2015, 268(1):328-339.
3.Lee AYS. A review of the role and clinical utility of anti-Ro52/TRIM21 in systemic autoimmunity[J]. Rheumatology International, 2017, 37(8):1323-1333.