Difference between revisions of "Part:BBa K3889022"
 
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<partinfo>BBa_K3889022 short</partinfo>
 
<partinfo>BBa_K3889022 short</partinfo>
  
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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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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 S99, S156 and S244 (amino acid numbered with reference to pro-ovastacin).
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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 S99, S156 and S244 (amino acid numbered with reference to pro-Ovastacin).
  
 
===Protein Sequence===
 
===Protein Sequence===
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RLLSAASNKWPMGGDGVVEVPFLLSSKYDEPSRQVILEALAEFERSTCIRFVTYQDQRDFISIIPMYGCFSDVGRSGGMQVVSLAPTCLQKGRGIVL<span style="color:red">HELMHVLGFWH</span>E HTRADRDRYIRVNWNEILPGFEINFIKSRSSNMLTPYDYSSVMHYGRLAFDRRGLPTITPLWAPSVHIGQRWNLSASDITRVLKLYGCS
 
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where <span style="color:red">red</span> denotes the active site [1]
 
 
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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">
 
<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
  
  

Latest revision as of 11:23, 19 October 2021


Human Ovastacin protease phosphomimic_A

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 S99, S156 and S244 (amino acid numbered with reference to pro-Ovastacin).

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