N-terminal cysteines essential for Golgi sorting of B-50 (GAP-43) in PC12 cells.
We have examined the subcellular distribution of the growth-associated protein B-50 (GAP-43) in pheochromocytoma (PC12) cells, using confocal microscopy. Proliferating PC12 cells contained very low levels of B-50 in the cytosol. Enhanced expression of B-50 in these cells, evoked by either nerve growth factor (NGF) treatment or transient transfection with rat B-50 cDNA, led to Golgi sorting and membrane targeting of the B-50 protein. Site directed mutagenesis of Cys3Cys4 to Ser3Gly4 in B-50 resulted in a cytosolic distribution. We conclude that Cys3, and Cys4 are essential for accumulation of B-50 both at the plasma membrane and in the Golgi apparatus of PC12 cells.
Aarts
L H
LH
Rudolf Magnus Institute for Neurosciences, Department of Physiological Chemistry, Utrecht University, The Netherlands.
van der Linden
J A
JA
Hage
W J
WJ
van Rozen
A J
AJ
Oestreicher
A B
AB
Gispen
W H
WH
Schotman
P
P
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D019922:GAP-43 Protein; D006056:Golgi Apparatus / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000378:metabolism*; D015345:Oligonucleotide Probes; D016716:PC12 Cells; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism
D018076:DNA, Complementary; D019922:GAP-43 Protein; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D015345:Oligonucleotide Probes; D003545:Cysteine