Purification and characterization of a calcium-activated neutral protease from monkey brain and its action on neuropeptides
- PMID: 6094523
- DOI: 10.1093/oxfordjournals.jbchem.a134895
Purification and characterization of a calcium-activated neutral protease from monkey brain and its action on neuropeptides
Abstract
A calcium-activated neutral protease was purified from Japanese monkey brain by ammonium sulfate fractionation and sequential column chromatographies monitored by assay of caseinolytic activity. The purified enzyme gave a single protein band on non-denaturing polyacrylamide gel electrophoresis, and consisted of two subunits with molecular weights of 74,000 and 20,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme required millimolar order calcium ions for activation, and was optimally active at pH 7.5-8.0. Upon incubation with various neuropeptides as substrates, the enzyme preferentially cleaved the peptide bonds with Arg, Lys, or Tyr at the P1 position and an amino acid residue with a bulky aliphatic side chain, such as Leu, Val, or Ile, at the P2 position. The hydrolytic activity toward neuropeptides as well as casein was strongly inhibited by various thiol protease inhibitors. These results suggested that the brain calcium-activated neutral protease may participate in the degradation of neuropeptides in vivo.
Similar articles
-
Purification and characterization of a calcium-activated neutral protease from monkey cardiac muscle.J Biochem. 1983 May;93(5):1435-45. doi: 10.1093/oxfordjournals.jbchem.a134279. J Biochem. 1983. PMID: 6309758
-
Degradation of neuropeptides by calcium-activated neutral protease.J Biochem. 1983 Dec;94(6):2071-4. doi: 10.1093/oxfordjournals.jbchem.a134564. J Biochem. 1983. PMID: 6323389
-
Purification and characterization of two forms of Ca2+-activated neutral protease from calf brain.J Biol Chem. 1983 Jul 25;258(14):8955-62. J Biol Chem. 1983. PMID: 6306006
-
Calcium-activated neutral protease (CANP) in brain and other tissues.Prog Neurobiol. 1984;23(1-2):63-78. doi: 10.1016/0301-0082(84)90012-1. Prog Neurobiol. 1984. PMID: 6097938 Review. No abstract available.
-
Characterization of neuropeptidases using inhibitors.Methods Mol Biol. 1997;73:369-81. doi: 10.1385/0-89603-399-6:369. Methods Mol Biol. 1997. PMID: 9031224 Review. No abstract available.
Cited by
-
Calcium-activated neutral protease activities in brain trauma.Neurochem Res. 1991 Apr;16(4):483-7. doi: 10.1007/BF00965570. Neurochem Res. 1991. PMID: 1922659
-
A simple one-step procedure for the separation of calpain I, calpain II and calpastatin.Biochem J. 1985 Oct 1;231(1):201-4. doi: 10.1042/bj2310201. Biochem J. 1985. PMID: 2998342 Free PMC article.
-
Molecular determinants of survival motor neuron (SMN) protein cleavage by the calcium-activated protease, calpain.PLoS One. 2010 Dec 30;5(12):e15769. doi: 10.1371/journal.pone.0015769. PLoS One. 2010. PMID: 21209906 Free PMC article.
-
Calmodulin-binding proteins as calpain substrates.Biochem J. 1989 Sep 15;262(3):693-706. doi: 10.1042/bj2620693. Biochem J. 1989. PMID: 2556106 Free PMC article. Review. No abstract available.
-
Calpain inhibition by peptide epoxides.Biochem J. 1985 Sep 1;230(2):509-16. doi: 10.1042/bj2300509. Biochem J. 1985. PMID: 2996503 Free PMC article.
