Proteomic Analysis of Fatty-acylated Proteins in Mammalian Cells with Chemical Reporters Reveals S-Acylation of Histone H3 Variants

Chemoproteomics reveals Toll-like receptor fatty acylation

“…After subtraction of proteins identified in the control lanes as described in the Methods Section and eliminating proteins with fewer than three spectral counts, a total of 247 proteins were identified from the DC2.4 samples and 437 proteins were identified from the MEF samples, indicating that there is a large difference in the repertoire of palmitoylated proteins expressed by these different cell types (see Additional file 1: Table S1, S2, Figure 1D). Of the proteins identified in DC2.4 cells, 76% had been previously identified as palmitoylated proteins in our past analysis of this cell line, and by others in proteomic studies of other cell types [7,16,17,[27][28][29][30][31] (see Additional file 1: Table S1). Of the proteins identified in MEFs, which had not been palmitoylome profiled before, 64% were previously reported to be palmitoylated, providing strong validation of our methodology (see Additional file 1: Table S2).…”

“…Likewise, the work identifying BST2 in B cells utilized acyl-biotin exchange chemistry, which allows specific enrichment of S-acylated proteins [16]. BST2 is also well characterized to be lipidated with a GPI anchor [39], which could incorporate long chain fatty acids such as alk-16, although previous alk-16 proteomic profiling studies have not been significantly contaminated with GPI anchored proteins, arguing against this possibility [7,9,11,17,27]. Mutation of individual residues within BST2 will be necessary to determine the precise location and type of lipidation reported by alk-16 labeling.…”

Linker for Activation of T-cell Family Member2 (LAT2) a Lipid Raft Adaptor Protein for AKT Signaling, Is an Early Mediator of Alkylphospholipid Anti-leukemic Activity

“…LAT2, RPL23, and CLIC1 contain sites of palmitoylation (37,38), a post-translational modification that is believed to target proteins to lipid rafts. Furthermore, when these six proteins were evaluated with respect to tissue expression using BioGPS (39), LAT2 was shown to be specifically expressed in hematopoietic tissues.…”

The Protein Lipidation and Its Analysis

“…The authors could prove with this approach that the palmitoyl turnover in Lck increases upon T-cell activation. Metabolic labeling with alkyne-containing fatty acids has also been used by other groups for the study the lipidated N-Ras and H-Ras, [127] for the characterization of myristoylated proteins [128] or for confirming the acylation of Histone H3 variants [129,130]. In addition, isoprenoid derivatives have also been reported and used for the analysis of the prenylome using CuAAC [116,117,131,132].…”