The development of mass spectrometry (MS)-based proteomics methods has been critical in providing new
insight about cellular processes and adaptations in all domains of life. While traditional MS-based methods
are not inherently quantitative, technologies are now available to overcome this limitation. Of note, stable
isotope labeling of amino acids in cell culture (SILAC) is reported as a reliable tool to label proteomes for
quantitative MS-based proteomics that is accurate and flexible for multiplexing. The isotopically labeled
lysine and arginine are easily incorporated into the proteome of cells auxotrophic for these amino acids.
Microorganisms of the domain Archaea provide a fascinating alternative to understanding cellular adaptations
and responses to environmental stresses. However, the availability of preferred SILAC-based quantitative
analyses is limited. This protocol describes the use of SILAC to quantitatively analyze the proteome of
Haloferax volcanii, a mesophilic halophilic archaeon that is easy to grow and requires no special equipment
to maintain.
Couto-Rodriguez, R.L., Gal, D., McMillan, L.J., Koh, J., Chen, S., Maupin-Furlow, J.A. (2022). Quantitative Mass Spectrometry by SILAC in Haloferax volcanii. In: Ferreira-Cerca, S. (eds) Archaea. Methods in Molecular Biology, vol 2522. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2445-6_16
Link to chapter: https://link.springer.com/protocol/10.1007/978-1-0716-2445-6_16