Distribution of calcium-activated neutral proteinase activity in quaking mouse brain: a subcellular study.
The distribution of calcium-activated neutral proteinase (CANP) activity was examined in the subcellular fractions of quaking and control mouse brain. The CANP activity was determined in purified myelin, cytosol and pellet (P2, consisting of nuclei, mitochondria and microsomes) fractions using [14C]azocasein as substrate. The enzyme activity in quaking brain was 1.3-fold greater than control. Fifty-seven percent of the control brain activity was in purified myelin compared to only 7% in quaking myelin. The specific activity of the control purified myelin was 4-fold greater than homogenate while that of the quaking was two-fold greater. In contrast, 51% of the quaking brain activity was present in cytosol compared to only 18% in the control. Triton X-100 greatly increased the control brain activity (10-fold) while the quaking brain activity was increased by only 1.2-fold. The total calcium content in the quaking brain was greatly elevated (6-fold) compared to control. Approximately 30% of the brain 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) activity was in quaking myelin while 77% of the CNPase activity in control brain was in myelin. These results suggest that in quaking brain much of the CANP is not incorporated into the myelin membrane and remains cytosolic.
Banik
N L
NL
Department of Neurology, Medical University of South Carolina, Charleston 29425.
Chakrabarti
A K
AK
Gantt
G
G
Hogan
E L
EL
D063309:2',3'-Cyclic Nucleotide 3'-Phosphodiesterase; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002154:Calpain / Q000302:isolation & purification / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology; D007700:Kinetics; D051379:Mice; D008821:Mice, Quaking / Q000378:metabolism*; D009186:Myelin Sheath / Q000201:enzymology; D010727:Phosphoric Diester Hydrolases; D012016:Reference Values; D013347:Subcellular Fractions / Q000201:enzymology
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D010727:Phosphoric Diester Hydrolases; D063309:2',3'-Cyclic Nucleotide 3'-Phosphodiesterase; C571338:Cnp protein, mouse; D002154:Calpain