Premium
Phosphorylation of Neurofilament Heavy‐Chain Side‐Arm Fragments by Cyclin‐Dependent Kinase‐5 and Glycogen Synthase Kinase‐3α in Transfected Cells
Author(s) -
Bajaj Narinder P. S.,
Miller Christopher C. J.
Publication year - 1997
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1997.69020737.x
Subject(s) - cyclin dependent kinase , gsk 3 , phosphorylation , kinase , microbiology and biotechnology , transfection , glycogen synthase , protein kinase domain , neurofilament , biology , cyclin dependent kinase 9 , gsk3b , biochemistry , chemistry , cyclin dependent kinase 2 , protein kinase a , cell , gene , cell cycle , immunohistochemistry , mutant , immunology
The side‐arm domain of neurofilament heavy‐chain (NF‐H) is heavily phosphorylated in axons. Much of this phosphate is located within a multiphosphorylation repeat (MPR) domain situated toward the carboxy terminus of the molecule. The MPR domain contains the repeat motif KSP of which there are two broad categories, KSPXX and KSPXK. In mouse NF‐H, the KSPXK repeats are situated toward the latter part of the MPR domain. We have expressed in mammalian cells fragments of mouse NF‐H side‐arm containing all of the MPR domain, the latter part of the MPR domain containing the KSPXK repeats, and the complementary amino‐terminal part of the MPR domain, which contains the KSPXX repeats. By cotransfecting these fragments with the neurofilament kinases cyclin‐dependent kinase‐5 (cdk‐5)/p35 and glycogen synthase kinase‐3α (GSK‐3α), we show that cdk‐5 induces cellular phosphorylation of the KSPXK‐containing fragment of NF‐H. Using the transfected fragments, we also map the epitopes for several commonly utilised NF‐H monoclonal antibodies and describe the effects that phosphorylation by cdk‐5 and GSK‐3α have on their reactivities.