Part:BBa_K4165015
UBE2W
Ubiquitin-conjugating E2 ligase that has a role in the ubiquitination cascade for protein degradation.
Usage and Biology
This E2 ubiquitin-conjugating enzyme UBE2W is the key participant in the set of enzymes as it is responsible for the initial step of monoubiquitylation by TRIM21 E3 ligase. The UBE2W is most specific for RING domain E3 ligases which happens to be that Trim21, which we are working with, is one of those RING domain E3 ligases.
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]
Dry Lab
Mathematical modeling
Transcription rate and translation rate under T7 promotor
the mathematical modeling was based on our code for the calculation of transcription and translation (you can find it in the code section) beside with the estimated results from the wet lab.
Figure 1. this figure shows the results from the transcription and translation code showing the
variation of mRNA and protein concentrations with time compared with the wet lab results.
WetLab Results
Transformation of His UBE 2w in BL-21 using pGS-21a vector
Figure 2. Transformed plate of His UBE 2w + pGS-21a
Transformation of His UBE 2w in DH-5 alpha using pJET vector
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Figure 3. Transformed plate of His UBE 2w + pJET
References
1. Stewart, M. D., Ritterhoff, T., Klevit, R. E., & Brzovic, P. S. (2016). E2 enzymes: more than just middle men. Cell research, 26(4), 423-440. 2. Vittal, V., Wenzel, D. M., Brzovic, P. S., & Klevit, R. E. (2013). Biochemical and structural characterization of the ubiquitin-conjugating enzyme UBE2W reveals the formation of a noncovalent homodimer. Cell biochemistry and biophysics, 67(1), 103-110.
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