A structural basis for substrate specificities of protein Ser/Thr kinases: primary sequence preference of casein kinases I and II, NIMA, phosphorylase kinase, calmodulin-dependent kinase II, CDK5, and Erk1.
We have developed a method to study the primary sequence specificities of protein kinases by using an oriented degenerate peptide library. We report here the substrate specificities of eight protein Ser/Thr kinases. All of the kinases studied selected distinct optimal substrates. The identified substrate specificities of these kinases, together with known crystal structures of protein kinase A, CDK2, Erk2, twitchin, and casein kinase I, provide a structural basis for the substrate recognition of protein Ser/Thr kinases. In particular, the specific selection of amino acids at the +1 and -3 positions to the substrate serine/threonine can be rationalized on the basis of sequences of protein kinases. The identification of optimal peptide substrates of CDK5, casein kinases I and II, NIMA, calmodulin-dependent kinases, Erk1, and phosphorylase kinase makes it possible to predict the potential in vivo targets of these kinases.
Songyang
Z
Z
Division of Signal Transduction, Beth Israel Hospital, Boston, Massachusetts 02215, USA.
Lu
K P
KP
Kwon
Y T
YT
Tsai
L H
LH
Filhol
O
O
Cochet
C
C
Brickey
D A
DA
Soderling
T R
TR
Bartleson
C
C
Graves
D J
DJ
DeMaggio
A J
AJ
Hoekstra
M F
MF
Blenis
J
J
Hunter
T
T
Cantley
L C
LC
D000595:Amino Acid Sequence; D042846:CDC2-CDC28 Kinases; D029742:Caenorhabditis elegans Proteins; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D047390:Casein Kinase II; D047388:Casein Kinases; D018797:Cell Cycle Proteins; D018360:Crystallography, X-Ray; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2; D051360:Cyclin-Dependent Kinase 5; D018844:Cyclin-Dependent Kinases / Q000737:chemistry / Q000378:metabolism; D016208:Databases, Factual; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D008958:Models, Molecular; D009124:Muscle Proteins / Q000737:chemistry / Q000378:metabolism; D000072380:NIMA-Related Kinase 1; D000072379:NIMA-Related Kinases; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010748:Phosphopeptides / Q000737:chemistry / Q000302:isolation & purification; D010764:Phosphorylase Kinase / Q000378:metabolism; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity
D029742:Caenorhabditis elegans Proteins; D002148:Calmodulin-Binding Proteins; D018797:Cell Cycle Proteins; D009124:Muscle Proteins; D009842:Oligopeptides; D010748:Phosphopeptides; C489233:unc-22 protein, C elegans; D011494:Protein Kinases; D010764:Phosphorylase Kinase; D047390:Casein Kinase II; D047388:Casein Kinases; D051360:Cyclin-Dependent Kinase 5; D000072380:NIMA-Related Kinase 1; D000072379:NIMA-Related Kinases; C442331:NIMA-related kinase 6; D017346:Protein Serine-Threonine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; C517679:unc-43 protein, C elegans; D042846:CDC2-CDC28 Kinases; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases