Purification and degradation of purified neurofilament proteins by the brain calcium-activated neutral proteases.
The effect of the three forms (CANP1, CANP2 & CANP3) of calf brain calcium activated neutral protease (CANP) on the hydrolysis of purified neurofilament triplet proteins was investigated. It was observed that: each of the purified neurofilament proteins, was hydrolyzed slowly by CANP2 whereas the hydrolysis of 150 KDa and 68 kDa proteins by CANP1 & CANP3 was rapid; when assembled neurofilaments were used as a substrate, again differences in the rate and extent of degradation of the triplet proteins by the three proteases were observed. For example, little cleavage of the 68 kDa protein by CANP2 and CANP3 was noted whereas 210 kDa and 150 kDa proteins remained largely intact. CANP1 degraded the 150 kDa and 68 kDa proteins more rapidly than 210 kDa protein, where only a slight effect was noted. These data provide further proof of the existence of three different forms of CANP in the brain, and indications of the resistance of 210 kDa protein to proteolysis which may be compatible with its proposed special role in crossbridge formation.
D000818:Animals; D001921:Brain / Q000201:enzymology*; D002118:Calcium / Q000502:physiology; D002417:Cattle; D002852:Chromatography, Ion Exchange; D003599:Cytoskeleton / Q000378:metabolism*; D010450:Endopeptidases / Q000502:physiology*; D066298:In Vitro Techniques; D007382:Intermediate Filaments / Q000378:metabolism*; D015260:Neprilysin; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism*
D009419:Nerve Tissue Proteins; D010450:Endopeptidases; D015260:Neprilysin; D002118:Calcium