Searching for substrates of lysosomal cathepsins

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For the discovery of substrates for cathepsin D, whole brain lysates from cathepsin D deficient (CD-/-) and the control littermate mice were separated by two-dimensional gel electrophoresis. Several proteins increased in CD-/- mouse brains were identified by mass spectrometry (MS). However immunohistochemical analyses revealed that some of the identified proteins were enriched in active microglia or astrocytes in CD-/- brains. We also found that the protein amount and activity of cathepsin C, whic h is a lysosomal protease and reported to be cleaved by cathepin D in vivo, was increased. However cathepsin C was also detected in activated microglia in CD-/- brains.