| Předmět: |
0301 basic medicine
chemistry.chemical_classification Growth medium 030102 biochemistry & molecular biology Brain Acid Soluble Protein 1 Organic Chemistry Fatty acid Myristic acid Biology medicine.disease_cause Biochemistry Lauric acid Yeast 03 medical and health sciences chemistry.chemical_compound chemistry medicine Molecular Medicine lipids (amino acids peptides and proteins) Molecular Biology Escherichia coli Myristoylation |
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| Zdroj: | ChemBioChem. |
| ISSN: | 1439-4227 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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