In this project, freshly isolated mitochondria from K-562 cells were crosslinked immediately with the homobifunctional and membrane-permeable reagent disuccinimidyl sulfoxide (DSSO). The study focuses on the comprehensive identification of proteins, specifically distinguishing between soluble and insoluble fractions. Both fractions were digested using Trypsin and analyzed using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS) and Data-Dependent Acquisition (DDA) methods. The resulting data were processed using MaxQuant software.