The aim of this project was to investigate the functional role of protein lactoacylation modification in mitochondrial metabolism. To achieve this, we isolated cellular mitochondria and conducted an immunoprecipitation (IP) assay using a pan-lactoacylation antibody (Pan-Kla) to analyze the lactoacylation proteome within mitochondria. Lactoacylation, a recently identified post-translational modification of proteins, has been shown to play a critical role in cellular metabolism and gene expression regulation. This study will employ mass spectrometry to identify and quantify the IP products, thereby elucidating the specific targets of lactoacylation modification in cancer cells and their biological significance. Our findings may provide novel potential targets for cancer therapy.