Global Profiling of Protein Lactylation Across Subcellular Fractionation Methods in HCT-116

ABSTRACT: Protein lactylation is a novel post-translational modification (PTM) involved in many important physiological processes such as macrophage polarization, immune regulation and tumour cell growth. However, most studies to date have focused on the function of lactylation on histones, and little is known about the distribution of lactylation on subcellular proteins. Here, we utilized subcellular fractionation methods to perform the global profiling of lactylation in human colon carcinoma HCT116 cells. This approach detected more unique lactylated proteins compared to the standard method when the amount of antibody and cells was equal. In total, 899 lysine lactylation sites were detected on 441 proteins, including 291 newly discovered lactylation sites and 63 newly lactylated proteins reported under the DDA and DIA acquisition modes. Functional enrichment analysis revealed that the majority of these proteins are involved in nucleosome assembly and slicesome function. For each subcellular fraction, the newly discovered lactylated proteins account for 10% to 20%. Notably, XPC not only has four new lactylation modification sites, but also one of which has been shown to be essential for the nucleotide excision repair process. Additionally, one of the newly discovered lactylation sites in scaffold attachment factor B1 (SAFB1) was found to be very important for the negative regulation of transcriptional activity. In conclusion, we describe a novel method that provides for understanding the characterization of subcellular lactylated proteins.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Colon

SUBMITTER: Bao qiuyu  

LAB HEAD: qiuyu Bao

PROVIDER: PXD053474 | Pride | 2024-12-15

REPOSITORIES: Pride