Phosphorylation and modulation of brain glutamate transporters by protein kinase C.
High affinity sodium- and potassium-coupled L-glutamate transport into presynaptic nerve terminals and fine glial processes removes the neurotransmitter from the synaptic cleft, thereby terminating glutamergic transmission. This report describes that the purified L-glutamate transporter from pig brain is phosphorylated by protein kinase C, predominantly at serine residues. Upon exposure of C6 cells, a cell line of glial origin, to 12-O-tetradecanoylphorbol-13-acetate, about a 2-fold stimulation of L-glutamate transport is observed within 30 min. Concomitantly, the level of phosphorylation increases with similar kinetics. The phorbol ester also stimulates L-glutamate transport in HeLa cells infected with a recombinant vaccinia virus expressing T7 RNA polymerase and transfected with pT7-GLT-1. The latter is a recently cloned rat brain glutamate transporter of glial origin. Mutation of serine 113 to asparagine does not affect the levels of expressed transport but abolishes its stimulation by the phorbol ester. To our knowledge, this is the first direct demonstration of the regulation of a neurotransmitter transporter by phosphorylation.
Casado
M
M
Centro de Biología Molecular, Facultad de Ciencias, Universidad Autónoma de Madrid, Spain.
Bendahan
A
A
Zafra
F
F
Danbolt
N C
NC
Aragón
C
C
Giménez
C
C
Kanner
B I
BI
D027322:Amino Acid Transport System X-AG; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D001921:Brain / Q000378:metabolism*; D002460:Cell Line; D017931:DNA Primers; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D006023:Glycoproteins / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D027322:Amino Acid Transport System X-AG; D017931:DNA Primers; D005971:Glutamates; D006023:Glycoproteins; D018698:Glutamic Acid; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate