Work

Metabolic Labeling of Ras with Tritiated Palmitate to Monitor Palmitoylation and Depalmitoylation

Year: 2013

Type: book-chapter

Abstract: Metabolic labeling with tritiated palmitate is a direct method for monitoring posttranslational modification of Ras proteins with this fatty acid. Advances in intensifying screens have allowed for the easy visualization of tritium without the need for extended exposure times. While more energetic radioisotopes are easier to visualize, the lack of commercial source and need for shielding make them more difficult to work with. Since radiolabeled palmitate is directly incorporated into Ras, its loss can be monitored by traditional pulse-chase experiments that cannot be accomplished with the method of acyl-exchange chemistry. As such, tritiated palmitate remains a readily accessible and direct method for monitoring the palmitoylation status of Ras proteins under a multitude of conditions.

Source: Methods in molecular biology

DOI: 10.1007/978-1-62703-791-4_3

Authors: Frederick D. Tsai, Joseph P. Wynne, Ian M. Ahearn, Mark R. Philips

Institutions: NYU Langone Health, New York University

Language: English

Topic: Protein Kinase Regulation and GTPase Signaling

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Cancer Institute, National Institute of General Medical Sciences

Awards: Compartment Specific Signaling of Ras, Regulation of K-Ras by a Farnesyl-electrostatic Switch, Medical Scientist Research Service Award