A calcium-activated neutral protease in the frog nervous system which degrades rapidly transported axonal proteins

A Ca2+-activated protease (CANP) was prepared from a soluble extract of the frog nervous system and separated from an endogenous high-molecular weight inhibitor by ion exchange chromatography. Its proteolytic action was tested on rapidly transported [3H]leucine-labelled proteins from sensory axons of the sciatic nerve. The enzyme was activated at neutral pH by Ca2+ at millimolar level, and was inhibited by leupeptin and the endogenous inhibitor. Its most pronounced degradative action was exerted towards two prominent rapidly transported peaks corresponding to 82 kDa and 90 kDa proteins. Proteolysis was accompanied by the appearance of mainly 16 kDa degradation products. When newly synthesized soluble proteins of the spinal dorsal ganglia were used as substrates, the presence of CANP resulted in degradation of proteins in a broad molecular-weight range (40-200 kDa).