N-Lauroylation during the Expression of Recombinant N-Myristoylated Proteins
- Author(s)
- Andrea Gabriele Flamm, Anabel-Lise Le Roux, Borja Mateos, Mireia Díaz-Lobo, Barbara Storch, Kathrin Breuker, Robert Konrat, Miquel Pons, Nicolas Coudevylle
- Abstract
Incorporation of myristic acid onto the N terminus of a protein is a crucial modification that promotes membrane binding and correct localization of important components of signaling pathways. Recombinant expression of N-myristoylated proteins in Escherichia coli can be achieved by co-expressing yeast N-myristoyltransferase and supplementing the growth medium with myristic acid. However, undesired incorporation of the 12-carbon fatty acid lauric acid can also occur (leading to heterogeneous samples), especially when the available carbon sources are scarce, as it is the case in minimal medium for the expression of isotopically enriched samples. By applying this method to the brain acid soluble protein 1 and the 1–185 N-terminal region of c-Src, we show the significant, and protein-specific, differences in the membrane binding properties of lauroylated and myristoylated forms. We also present a robust strategy for obtaining lauryl-free samples of myristoylated proteins in both rich and minimal media.
- Organisation(s)
- Department of Structural and Computational Biology
- External organisation(s)
- Universitat Autònoma de Barcelona , Institute for Research in Biomedicine, Leopold-Franzens-Universität Innsbruck, Max F. Perutz Laboratories GmbH (MFPL)
- Journal
- ChemBioChem: a european journal of chemical biology
- Volume
- 17
- Pages
- 82-89
- No. of pages
- 8
- ISSN
- 1439-4227
- DOI
- https://doi.org/10.1002/cbic.201500454
- Publication date
- 12-2015
- Peer reviewed
- Yes
- Austrian Fields of Science 2012
- 106041 Structural biology
- Keywords
- ASJC Scopus subject areas
- Molecular Medicine, Molecular Biology, Biochemistry, Organic Chemistry
- Portal url
- https://ucris.univie.ac.at/portal/en/publications/nlauroylation-during-the-expression-of-recombinant-nmyristoylated-proteins(04f746a1-cd14-4cb4-985d-d08e6a5f1065).html