Premium
Calcium‐dependent association of annexin VI, protein kinase Cα, and neurocalcinα on the raft fraction derived from the synaptic plasma membrane of rat brain
Author(s) -
Orito Aya,
Kumanogoh Haruko,
Yasaka Kaoru,
Sokawa Junko,
Hidaka Hiroyoshi,
Sokawa Yoshihiro,
Maekawa Shohei
Publication year - 2001
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.1071
Subject(s) - raft , lipid raft , lipid microdomain , biochemistry , protein kinase c , calcium binding protein , calmodulin , biology , annexin , protein kinase a , membrane protein , microbiology and biotechnology , chemistry , kinase , signal transduction , calcium , membrane , enzyme , organic chemistry , cell , copolymer , polymer
A membrane microdomain enriched in cholesterol and sphingolipids or so called “raft” region was found to contain many signal transducing proteins such as GPI‐anchored proteins, trimeric G proteins and protein tyrosine kinases. Because brain‐derived raft contains two calmodulin‐binding proteins, GAP‐43 and NAP‐22 as the major protein components, the raft domain is assumed to be important in the Ca 2+ ‐signaling. In this study, we analyzed protein components showing Ca 2+ ‐dependent binding to the raft of synaptic plasma membrane from rat brain. SDS‐PAGE analysis of the protein components in the EGTA eluate from the raft prepared in the presence of Ca 2+ ‐ions showed the elution of 80 kDa, 68 kDa, 22 kDa, and 21 kDa proteins. These proteins were identified as protein kinase Cα (80 kDa) and annexin VI (68 kDa) from the partial amino‐acid sequencing, and neurocalcinα (22 kDa) and calmodulin (21 kDa) with western blotting and electrophoretic mobilities in the presence or absence of Ca 2+ ions. Further immunoblotting experiments showed the Ca 2+ ‐dependent association of conventional, but not non‐conventional, subtypes of PKC to the raft. J. Neurosci. Res. 64:235–241, 2001. © 2001 Wiley‐Liss, Inc.