title: Calcium-activated neutral proteinase in rat brain myelin and subcellular fractions.
abstract: The activity of calcium-activated neutral proteinase (mM CANP) was determined in subcellular fractions of rat brain. The CANP activity in whole homogenate and its membrane fractions including myelin was increased ten-fold following treatment with Triton X-100. The majority of the activity (60%) was in the primary particulate fractions P1 (nuclear), P2 (mitochondrial), and P3 (microsomal). Following subfractionation of each particulate fraction, most of the activity (50%) was found in the myelin-enriched fractions (P1A, P2A, and P3A) and separated at the interface of 0.32-0.85 M sucrose. Only 20-30% of the total homogenate activity was in cytosol. The enrichment in the myelin fractions resembled that for 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) activity. Immunoblotting revealed that the CANP was mainly in myelin and cytosol. In addition to the presence of 72-76 Kd and 80 Kd bands, there were faint high-molecular-weight CANP bands ranging from 110-150 Kd and lower-molecular-weight forms in the region of 30-50 Kd in both purified myelin and cytosol. These studies suggested that CANP is present in myelin and cytosol and that it exists in the brain in membrane-bound and soluble forms.
authors.affiliation: Department of Neurology, Medical University of South Carolina, Charleston 29425.
authors.forename: A K
authors.initials: AK
authors.lastname: Chakrabarti
authors.forename: Y
authors.initials: Y
authors.lastname: Yoshida
authors.forename: J M
authors.initials: JM
authors.lastname: Powers
authors.forename: I
authors.initials: I
authors.lastname: Singh
authors.forename: E L
authors.initials: EL
authors.lastname: Hogan
authors.forename: N L
authors.initials: NL
authors.lastname: Banik
mesh_terms: D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000032:analysis; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002154:Calpain / Q000032:analysis*; D008297:Male; D009186:Myelin Sheath / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D002154:Calpain