A novel protein phosphatase that dephosphorylates and regulates Ca2+/calmodulin-dependent protein kinase II.

The Journal of Biological Chemistry
A IshidaH Fujisawa

Abstract

A synthetic peptide corresponding to the autophosphorylation site of Ca2+/calmodulin-dependent protein kinase II (CaMKII) (residues 281-289) was conjugated to paramagnetic particles, and phosphorylated by a constitutively active CaMKII fragment. Using this phosphopeptide conjugate as a substrate, a calyculin A-insensitive, Mn(2+)-dependent, and poly-L-lysine-stimulated protein phosphatase activity was detected in the crude extract of rat brain. The protein phosphatase (designated as CaMKII phosphatase) (CaMKIIPase) was purified to near homogeneity from rat brain. CaMKIIPase showed apparent molecular weights of 54,000 and 65,000, on SDS-polyacrylamide gel electrophoresis and gel-filtration analysis, respectively. It was not inhibited by 100 nM calyculin A or 10 microM okadaic acid. Mn2+, but not Mg2+, was absolutely required for activity. CaMKIIPase was potently activated by polycations. Autophosphorylated CaMKII was dephosphorylated by CaMKIIPase, whereas phosphorylase kinase, mixed histones, myelin basic protein, and alpha-casein (which had been phosphorylated by cAMP-dependent protein kinase) and phosphorylase a (phosphorylated by phosphorylase kinase) were not significantly dephosphorylated. No other proteins than CaMKII in ...Continue Reading

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Citations

Dec 5, 2002·Archives of Biochemistry and Biophysics·Atsuhiko IshidaHitoshi Fujisawa
Dec 17, 2005·Nature Immunology·Elena M GalloGerald R Crabtree
Sep 21, 2012·Journal of Biochemistry·Hiromi BabaIsamu Kameshita
Mar 27, 2001·Annual Review of Pharmacology and Toxicology·S S Hook, A R Means
Sep 28, 2007·Biochemical and Biophysical Research Communications·Noriyuki SueyoshiAtsuhiko Ishida
Aug 10, 2015·Molecular and Cellular Neurosciences·Johanna G PasekRoger J Colbran
Jan 5, 2010·Analytical Biochemistry·Isamu KameshitaNoriyuki Sueyoshi
Nov 6, 2013·Biochimica Et Biophysica Acta·Martin W Berchtold, Antonio Villalobo
Aug 28, 2016·Analytical Biochemistry·Yasunori SugiyamaIsamu Kameshita
Dec 16, 2016·Journal of Proteome Research·Songjing ZhangCheng-Gee Koh
Dec 11, 2007·Critical Reviews in Biochemistry and Molecular Biology·Twan Lammers, Sara Lavi
Jan 17, 2020·Cell Cycle·Pu Rum KimCheng-Gee Koh
Mar 14, 2019·Oncotarget·Muhammad Bakhait RahmatCheng-Gee Koh
Sep 23, 2003·The Journal of Immunology : Official Journal of the American Association of Immunologists·Qian ZhanJohn A Badwey
Mar 15, 2008·The Biochemical Journal·Samudra S GangopadhyayKathleen G Morgan

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