Difference between revisions of "Part:BBa K2271103"
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K2271103 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2271103 SequenceAndFeatures</partinfo> | ||
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+ | ===References=== | ||
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+ | [1] <b> Targeting of the tail-anchored peroxisomal membrane proteins PEX26 and PEX15 occurs through C-terminal PEX19-binding sites. </b> <br> | ||
+ | André Halbach, Christiane Landgraf, Stephan Lorenzen, Katja Rosenkranz, Rudolf Volkmer-Engert, Ralf Erdmann, and Hanspeter Rottensteiner (2006) <br> | ||
+ | <i>Journal of Cell Science 119, 2508-2517</i> Published by The Company of Biologists 2006 doi:10.1242/jcs.02979 | ||
+ | |||
+ | [2] <b> Overexpression of Pex15p, a phosphorylated peroxisomal integral membrane protein required for peroxisome assembly in <i>S.cerevisiae</i>, causes proliferation of the endoplasmic reticulum membrane. </b> <br> | ||
+ | Ype Elgersma, Liane Kwast, Marlene van den Berg, William B. Snyder, Ben Distel, Suresh Subramani, Henk F. Tabak(1997) <br> | ||
+ | <i>EMBO J 16(24):7326-41</i> DOI 10.1093/emboj/16.24.7326 | ||
Latest revision as of 22:29, 31 October 2017
pex15
Biology and Usage
This part contains a truncated version of Pex15 (aa315-383) from S. cerevisiea . [1]
Pex15 is a tail anchord peroxisomal membrane protein with its c-terminus directed to the peroxisomal lumen and its n-terminus directed to the cytosol.
It is required for peroxisome biogenesis. In cells lacking Pex15 peroxisomal matrix proteins mislocalize to the cytosol while overexpression results in impaired peroxisome assembly. [2]
Experimental design
The functionality of this part was validated by microscopy. For the microscopy mVenus was fused to the C-Terminus of Pex15. We could observe a peroxisomal typical localisation in the cell.
Additionaly we tested this part as a membrane anchor for the v-SNARE Snc1. We co-expressed the Snc1-Pex15 construct with GUS-PTS1 to perform a GUS Assay. The secreted GUS in the supernatant was measured with the turnover of 4-methylumbelliferyl-beta-D-glucuronide to 4-methyl umbelliferone (4-MU). The fluorescent 4-MU was measured with a plate reader (excitation: 365 nm, emission: 465 nm).
![](/wiki/images/a/ac/Igemduscgn2017GUSassay.png)
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] Targeting of the tail-anchored peroxisomal membrane proteins PEX26 and PEX15 occurs through C-terminal PEX19-binding sites.
André Halbach, Christiane Landgraf, Stephan Lorenzen, Katja Rosenkranz, Rudolf Volkmer-Engert, Ralf Erdmann, and Hanspeter Rottensteiner (2006)
Journal of Cell Science 119, 2508-2517 Published by The Company of Biologists 2006 doi:10.1242/jcs.02979
[2] Overexpression of Pex15p, a phosphorylated peroxisomal integral membrane protein required for peroxisome assembly in S.cerevisiae, causes proliferation of the endoplasmic reticulum membrane.
Ype Elgersma, Liane Kwast, Marlene van den Berg, William B. Snyder, Ben Distel, Suresh Subramani, Henk F. Tabak(1997)
EMBO J 16(24):7326-41 DOI 10.1093/emboj/16.24.7326