TY - JOUR
T1 - Phosphorylation site specificities of glycogen synthase kinases
T2 - Determination by peptide mapping using high-performance liquid chromatography
AU - Juhl, Henning
AU - Sheorain, Virender S.
AU - Schworer, Charles M.
AU - Jett, Mary Frances
AU - Soderling, Thomas R.
N1 - Funding Information:
‘Supported in part hy NIH grant AM17808. fellowship AM06449 to C.S., and a special fellowship to H. J. by the Danish Medical Research Council. ‘Present address: Department of Medicine, Mar-selisborg Hospital, P. P. Orumsgade 11, 8000 Arhus, Denmark. 3Author to whom correspondence should be addressed.
PY - 1983/4/15
Y1 - 1983/4/15
N2 - A method is described which separates the various phosphorylation sites in glycogen synthase based on reverse phase high-performance liquid chromatography (HPLC) of tryptic 32P-peptides. Using this method we studied the phosphorylation site specificities of the kinases which act on glycogen synthase. The cAMP-dependent protein kinase phosphorylated sites 1a, 1b, and 2, whereas casein kinase II phosphorylated only site 5. Two calcium, calmodulin-dependent kinases, phosphorylase kinase and liver calmodulin-dependent synthase kinase, both phosphorylated site 2, and the latter enzyme also phosphorylated site 1b. A cAMP-independent kinase (kinase 4) purified from liver also specifically phosphorylated site 2. Synthase kinase 3 catalyzed the phosphorylation of only site 3. This HPLC method was also used to establish that all of these sites were subject to phosphorylation in vivo.
AB - A method is described which separates the various phosphorylation sites in glycogen synthase based on reverse phase high-performance liquid chromatography (HPLC) of tryptic 32P-peptides. Using this method we studied the phosphorylation site specificities of the kinases which act on glycogen synthase. The cAMP-dependent protein kinase phosphorylated sites 1a, 1b, and 2, whereas casein kinase II phosphorylated only site 5. Two calcium, calmodulin-dependent kinases, phosphorylase kinase and liver calmodulin-dependent synthase kinase, both phosphorylated site 2, and the latter enzyme also phosphorylated site 1b. A cAMP-independent kinase (kinase 4) purified from liver also specifically phosphorylated site 2. Synthase kinase 3 catalyzed the phosphorylation of only site 3. This HPLC method was also used to establish that all of these sites were subject to phosphorylation in vivo.
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U2 - 10.1016/0003-9861(83)90550-7
DO - 10.1016/0003-9861(83)90550-7
M3 - Article
C2 - 6303223
AN - SCOPUS:0021103856
SN - 0003-9861
VL - 222
SP - 518
EP - 526
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 2
ER -