Difference between revisions of "Part:BBa K3889022"
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<partinfo>BBa_K3889022 short</partinfo> | <partinfo>BBa_K3889022 short</partinfo> | ||
− | + | 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. | |
+ | 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=== | ||
+ | <html> | ||
+ | <head> | ||
+ | <style> | ||
+ | div.scroll {overflow-x: aut0; overflow-y: hidden;} | ||
+ | </style> | ||
+ | </head> | ||
+ | <body> | ||
+ | <div class="scroll"> | ||
+ | <p>RLLSAASNKWPMGGDGVVEVPFLLSSKYDEPSRQVILEALAEFERSTCIRFVTYQDQRDFISIIPMYGCFSDVGRSGGMQVVSLAPTCLQKGRGIVL<span style="color:red">HELMHVLGFWH</span>EHTRADRDRYIRVNWNEILPGFEINFIKSRSSNMLTPYDYSSVMHYGRLAFDRRGLPTITPLWAPSVHIGQRWNLSASDITRVLKLYGCS</p> | ||
+ | </div> | ||
+ | <p>where <span style="color:red">red</span> denotes the active site [1]</p> | ||
+ | <div class="scroll"> | ||
+ | <img src="https://static.igem.org/mediawiki/parts/8/8e/T--IISER-Tirupati_India--ovastacin_allignment.jpg"> | ||
+ | </div> | ||
+ | </body> | ||
+ | </html> | ||
+ | Fig source: Geneious version 2021.2 created by Biomatters. Available from https://www.geneious.com | ||
+ | |||
+ | |||
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<partinfo>BBa_K3889022 parameters</partinfo> | <partinfo>BBa_K3889022 parameters</partinfo> | ||
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+ | |||
+ | |||
+ | ===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 |
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]
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]
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