Phosphorylation site specificities of glycogen synthase kinases: Determination by peptide mapping using high-performance liquid chromatography

Henning Juhl; Virender S. Sheorain; Charles M. Schworer; Mary Frances Jett; Thomas R. Soderling

doi:10.1016/0003-9861(83)90550-7

Phosphorylation site specificities of glycogen synthase kinases: Determination by peptide mapping using high-performance liquid chromatography

Henning Juhl
, Virender S. Sheorain
, Charles M. Schworer
, Mary Frances Jett
, Thomas R. Soderling

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	518-526
Number of pages	9
Journal	Archives of Biochemistry and Biophysics
Volume	222
Issue number	2
DOIs	https://doi.org/10.1016/0003-9861(83)90550-7
State	Published - Apr 15 1983
Externally published	Yes

Funding

‘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.

Funders	Funder number
Author National Institutes of Health National Institutes of Health National Institutes of Health National Institutes of Health The Bev Hartig Huntington's Disease Foundation National Institutes of Health	AM06449, AM17808
Sundhed og Sygdom, Det Frie Forskningsråd

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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10.1016/0003-9861(83)90550-7

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@article{604fed5ebff74375aa549a2383ea81c9,

title = "Phosphorylation site specificities of glycogen synthase kinases: Determination by peptide mapping using high-performance liquid chromatography",

abstract = "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.",

author = "Henning Juhl and Sheorain, \{Virender S.\} and Schworer, \{Charles M.\} and Jett, \{Mary Frances\} and Soderling, \{Thomas R.\}",

note = "Funding Information: {\textquoteleft}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. {\textquoteleft}Present address: Department of Medicine, Mar-selisborg Hospital, P. P. Orumsgade 11, 8000 Arhus, Denmark. 3Author to whom correspondence should be addressed.",

year = "1983",

month = apr,

day = "15",

doi = "10.1016/0003-9861(83)90550-7",

language = "English (US)",

volume = "222",

pages = "518--526",

journal = "Archives of Biochemistry and Biophysics",

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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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AN - SCOPUS:0021103856

SN - 0003-9861

VL - 222

SP - 518

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JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

IS - 2

ER -

Phosphorylation site specificities of glycogen synthase kinases: Determination by peptide mapping using high-performance liquid chromatography

Abstract

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ASJC Scopus subject areas

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