The selective degradation of injected proteins occurs principally in the cytosol rather than in lysosomes.

These studies use microinjection to determine whether the selective degradation of cytosolic proteins involves selective transfer of proteins to lysosomes or selective proteolysis within the cytosol. 14C-Sucrose-labeled bovine serum albumin (14C-sucBSA) was conjugated to polylysine, and monolayers of L929 cells were exposed to the conjugate. The 14C-sucrose-labeled peptides that arose upon degradation of the added 14C-sucBSA polylysine accumulated exclusively within lysosomes. In contrast, when 14C-sucBSA or 14C-sucrose-labeled pyruvate kinase (14C-sucPK) was microinjected into L929 cells, over half the 14C-sucrose-labeled peptides derived form the injected proteins were present in the postlysosomal supernatant. Control experiments demonstrated that the microinjection procedure did not cause 14C-sucrose peptides to leak from lysosomes. Therefore, the presence in the cytosol of substantial amounts of the degradation products from injected 14C-sucBSA and 14C-sucPK confirms the existence of a major proteolytic system(s) within or readily accessible to the cytosol of animal cells.