Phosphorylation of glial fibrillary acidic protein and vimentin by cytoskeletal-associated intermediate filament protein kinase activity in astrocytes.
These studies describe a cytoskeletal-associated protein kinase activity in astrocytes that phosphorylated the intermediate filament proteins glial fibrillary acidic protein (GFAP) and vimentin and that appeared to be distinct from protein kinase C (PK-C) and the cyclic AMP-dependent protein kinase (PK-A). The cytoskeletal-associated kinase activity phosphorylated intermediate filament proteins in the presence of 10 mM MgCl2 and produced an even greater increase in 32P incorporation into these proteins in the presence of calcium/calmodulin. Tryptic peptide mapping of phosphorylated intermediate filament proteins showed that the intermediate filament protein kinase activity produced unique phosphopeptide maps, in both the presence and the absence of calcium/calmodulin, as compared to that of PK-C and PK-A, although there were some common sites of phosphorylation among the kinases. In addition, it was determined that the intermediate filament protein kinase activity phosphorylated both serine and threonine residues of the intermediate filament proteins, vimentin and GFAP. However, the relative proportion of serine and threonine residues phosphorylated varied depending on the presence or absence of calcium/calmodulin. The magnesium-dependent activity produced the highest proportion of threonine phosphorylation, suggesting that the calcium/calmodulin-dependent kinase activity acts mainly at serine residues. PK-A and PK-C phosphorylated mainly serine residues. Also, the intermediate filament protein kinase activity phosphorylated both the N-and the C-terminal domains of vimentin and the N-terminal domain of GFAP. In contrast, both PK-C and PK-A are known to phosphorylate the N-terminal domains of both proteins.(ABSTRACT TRUNCATED AT 250 WORDS)
Harrison
B C
BC
Department of Pharmacology, University of Texas Health Science Center, San Antonio 78284.
Mobley
P L
PL
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001253:Astrocytes / Q000201:enzymology*; D002122:Calcium Chloride / Q000494:pharmacology; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000201:enzymology*; D005904:Glial Fibrillary Acidic Protein / Q000737:chemistry / Q000378:metabolism*; D007382:Intermediate Filaments / Q000201:enzymology*; D015636:Magnesium Chloride / Q000494:pharmacology; D010449:Peptide Mapping; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D014746:Vimentin / Q000737:chemistry / Q000378:metabolism*
D000596:Amino Acids; D005904:Glial Fibrillary Acidic Protein; D014746:Vimentin; D015636:Magnesium Chloride; D011494:Protein Kinases; D002122:Calcium Chloride