Difference between revisions of "Part:BBa K3889023"

 
 
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<partinfo>BBa_K3889023 short</partinfo>
 
<partinfo>BBa_K3889023 short</partinfo>
  
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Phosphomimiced form of Human Ovastacin protease with serine residues phosphomimetic for prokaryotic expression.
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Since Ovastacin (<partinfo>BBa_K3889024</partinfo>) has a eukaryotic origin, this protein undergoes several Post Translational Modifications (PTMs) including the addition of disulphide bonds and phosphorylation at serine and tyrosine residues.
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However, the recombinant protein if produced in a prokaryotic organism will lack the necessary machinery to carry out PTMs. Hence, a phosphomimetic variant(s) of Ovastacin was made by changing or mutating the serine to aspartic acid and tyrosine to glutamic acid residues at S99, S156, S244 and Y279 (amino acid numbered with reference to pro-ovastacin) respectively.
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===Protein Sequence===
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<p>RLLSAASNKWPMGGDGVVEVPFLLSSKYDEPSRQVILEALAEFERSTCIRFVTYQDQRDFISIIPMYGCFSDVGRSGGMQVVSLAPTCLQKGRGIVL<span style="color:red">HELMHVLGFWH</span>EHTRADRDRYIRVNWNEILPGFEINFIKSRSSNMLTPYDYSSVMHYGRLAFDRRGLPTITPLWAPSVHIGQRWNLSASDITRVLKLYGCS</p>
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<p>where <span style="color:red">red</span> denotes the active site [1]</p>
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<img src="https://static.igem.org/mediawiki/parts/8/8e/T--IISER-Tirupati_India--ovastacin_allignment.jpg">
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Fig source: Geneious version 2021.2 created by Biomatters. Available from https://www.geneious.com
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<partinfo>BBa_K3889023 parameters</partinfo>
 
<partinfo>BBa_K3889023 parameters</partinfo>
 
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===References===
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1. Anna D. Burkart, Bo Xiong, Boris Baibakov, Maria Jiménez-Movilla, Jurrien Dean; Ovastacin, a cortical granule protease, cleaves ZP2 in the zona pellucida to prevent polyspermy. J Cell Biol 2 April 2012; 197 (1): 37–44. doi: https://doi.org/10.1083/jcb.201112094

Latest revision as of 23:06, 19 October 2021


Human Ovastacin protease phosphomimic_B

Since Ovastacin (BBa_K3889024) has a eukaryotic origin, this protein undergoes several Post Translational Modifications (PTMs) including the addition of disulphide bonds and phosphorylation at serine and tyrosine residues. However, the recombinant protein if produced in a prokaryotic organism will lack the necessary machinery to carry out PTMs. Hence, a phosphomimetic variant(s) of Ovastacin was made by changing or mutating the serine to aspartic acid and tyrosine to glutamic acid residues at S99, S156, S244 and Y279 (amino acid numbered with reference to pro-ovastacin) respectively.

Protein Sequence

RLLSAASNKWPMGGDGVVEVPFLLSSKYDEPSRQVILEALAEFERSTCIRFVTYQDQRDFISIIPMYGCFSDVGRSGGMQVVSLAPTCLQKGRGIVLHELMHVLGFWHEHTRADRDRYIRVNWNEILPGFEINFIKSRSSNMLTPYDYSSVMHYGRLAFDRRGLPTITPLWAPSVHIGQRWNLSASDITRVLKLYGCS

where red denotes the active site [1]

Fig source: Geneious version 2021.2 created by Biomatters. Available from https://www.geneious.com


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



References

1. Anna D. Burkart, Bo Xiong, Boris Baibakov, Maria Jiménez-Movilla, Jurrien Dean; Ovastacin, a cortical granule protease, cleaves ZP2 in the zona pellucida to prevent polyspermy. J Cell Biol 2 April 2012; 197 (1): 37–44. doi: https://doi.org/10.1083/jcb.201112094