Macroautophagy is a degradation process characterized by the formation of double-membrane vesicles-autophagosomes that deliver cytoplasm and sequestered cargo to the lysosome.We have previously shown that several RabGAP proteins bind to a small ubiquitin modifiers ATG8,that are essential for the formation of autophagosomes and selective removal of cargo.We hypothesized that study of RabGAP proteins will lead us to a better conceptual understanding of autophagy, considering close relation of autophagy to other membrane trafficking events.Here, we show that previously reported TBC1D2B, which binds LC3, can regulate autophagy via Atg5/Atg16 complex and early endosomal GTPases.Using SILAC based mass spectrometry approach we identified novel binding partners of TBC1D2B.We provide the novel data on TBC1D2B function in endocytosis and autophagy in mouse embryonic fibroblasts (MEFs) isolated from TBC1D2B wild type and knock-out mice.TBC1D2B deficiency leads to a defect in autophagy and we show that both, binding to LC3 and to early endosomes are required for efficient rescue of the phenotype.