Difference between revisions of "Part:BBa K2271060"
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Gerst, Jeffrey E. (1997) <br> | Gerst, Jeffrey E. (1997) <br> | ||
<i>J. Biol. Chem. 272 (26), pp. 16591–16598.</i> DOI: 10.1074/jbc.272.26.16591. | <i>J. Biol. Chem. 272 (26), pp. 16591–16598.</i> DOI: 10.1074/jbc.272.26.16591. | ||
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+ | [2] <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) | ||
+ | In Journal of Cell Science 119, 2508-2517 Published by The Company of Biologists 2006 doi:10.1242/jcs.02979 | ||
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Revision as of 22:39, 1 November 2017
Snc1
Usage and Biology
![](/wiki/images/5/51/Snc1duscgn.png)
This part is a truncated version of the v-SNARE (vesicle- synaptosome-associated-Soluble N-ethylmaleimide-sensitive-factor Attachment REceptorprotein) Snc1. Snc1 is in the wildtype form involved in the fusion of Golgi-derived secretory vesicles with the plasma membrane. Domains of the protein are a variable domain which is not important for the binding to the t-SNARE, H1 and H2 are the α-helical segments (forming the SNAREpin with the t-SNARE) and the transmembrane domain.[1] For our approaches we used a truncated version without the transmembrane domain. This Snc1 truncation was fused to the N-Terminus of different peroxisomal membrane anchor (Pex15 /PEX26) to secrete the compounds of this compartment.
![](/wiki/images/d/d1/T--Cologne-Duesseldorf--SecretionSNAREpic.png)
Experimental design
For testing this part we used a fusion with the N-terminus of a peroxisomal membrane anchor. We co-expressed this construct with GUS-PTS1 (beta-glucuronidase)to perform a [http://2017.igem.org/Team:Cologne-Duesseldorf/Experiments 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).
Results
Different peroxisomal membrane anchors were tested using the GUS-assay. The highest activity of GUS could be measured in the supernatant of Pex15 as a membrane anchor.
![](/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] Conserved α-Helical Segments on Yeast Homologs of the Synaptobrevin/VAMP Family of v-SNAREs Mediate Exocytic Function.
Gerst, Jeffrey E. (1997)
J. Biol. Chem. 272 (26), pp. 16591–16598. DOI: 10.1074/jbc.272.26.16591.
[2] 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)
In Journal of Cell Science 119, 2508-2517 Published by The Company of Biologists 2006 doi:10.1242/jcs.02979