Phosphorylation of the beta-amyloid precursor protein at the cell surface by ectocasein kinases 1 and 2.
The beta-amyloid precursor protein (betaAPP) is one of the rare proteins known to be phosphorylated within its ectodomain. We have shown previously that betaAPP can be phosphorylated within secretory vesicles and at the cell surface (Walter, J., Capell, A., Hung, A. Y. , Langen, H., Schnölzer, M., Thinakaran, G., Sisodia, S. S., Selkoe, D. J., and Haass, C. (1997) J. Biol. Chem. 272, 1896-1903). We have now specifically characterized the phosphorylation of cell surface-located betaAPP and identified two ectoprotein kinases that phosphorylate betaAPP at the outer face of the plasma membrane. By using selective protein kinase inhibitors and by investigating the usage of ATP and GTP as cosubstrates, we demonstrate that membrane-bound betaAPP as well as secreted forms of betaAPP can be phosphorylated by casein kinase (CK) 1- and CK2-like ectoprotein kinases. The ectodomain of betaAPP was also phosphorylated by purified CK1 and CK2 in vitro, but not by protein kinases A and C. Phosphorylation of betaAPP by ectoprotein kinases and by purified CK1 and CK2 occurred within an acidic domain in the N-terminal half of the protein. Heparin strongly inhibited the phosphorylation of cell-surface betaAPP by ecto-CK1 and ecto-CK2, indicating a regulatory role of this extracellular matrix component in betaAPP phosphorylation.
Walter
J
J
Adolf-Butenandt-Institut, Department of Biochemistry, Laboratory for Alzheimer's Disease Research, Ludwig-Maximilians-Universät München, Schillerstrasse 44, D-80336 Munich, Germany. jwalter@pbm.med.uni-muenchen.de
Schindzielorz
A
A
Hartung
B
B
Haass
C
C
D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D047390:Casein Kinase II; D047388:Casein Kinases; D002462:Cell Membrane / Q000378:metabolism*; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D008954:Models, Biological; D010766:Phosphorylation / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D017434:Protein Structure, Tertiary
D016564:Amyloid beta-Protein Precursor; D006493:Heparin; D011494:Protein Kinases; C034454:ectoprotein kinase; D047390:Casein Kinase II; D047388:Casein Kinases; D017346:Protein Serine-Threonine Kinases