[29] Use of peptide substrates to study the specificity of phosphorylase kinase phosphorylation

1983 ◽  
pp. 268-278 ◽  
Author(s):  
Donald J. Graves
Keyword(s):  
Phosphorylase Kinase ◽  
Peptide Substrates ◽  
Kinase Phosphorylation

scholarly journals Studies on the specificity of phosphorylase kinase using peptide substrates.

1977 ◽  
Vol 252 (16) ◽  
pp. 5666-5671 ◽  
Author(s):  
G W Tessmer ◽  
J R Skuster ◽  
L B Tabatabai ◽  
D J Graves
Keyword(s):  
Phosphorylase Kinase ◽  
Peptide Substrates

scholarly journals Phosphorylase kinase phosphorylation of skeletal-muscle troponin T

1980 ◽  
Vol 191 (3) ◽  
pp. 851-854 ◽  
Author(s):  
V V Risnik ◽  
A B Dobrovolskii ◽  
N B Gusev ◽  
S E Severin
Keyword(s):  
Skeletal Muscle ◽  
Amino Acid ◽  
Troponin T ◽  
Rabbit Skeletal Muscle ◽  
Phosphorylase Kinase ◽  
Amino Acid Residues ◽  
Standard Preparation ◽  
The Third ◽  
Original Preparation ◽  
Kinase Phosphorylation

Rabbit skeletal-muscle troponin T was phosphorylated by a standard preparation of phosphorylase kinase [Cohen (1973) Eur. J. Biochem. 34, 1–14] and by fractions obtained after chromatography of phosphorylase kinase on phosphocellulose. The original preparation of phosphorylase kinase phosphorylated at least two sites, one of which was serine-1. The second and probably the third sites were presumably located in the peptide flanked by amino-acid residues 147 and 161 of troponin T. Fractions of phosphorylase kinase was adsorbed on phosphocellulose phosphorylated only the second site. Tightly adsorbed fractions possessed high troponin T kinase and phosvitin kinase activities and phosphorylated only serine-1 of troponin T. The results suggest that standard preparations of phosphorylase kinase are contaminated by troponin T kinase, which can phosphorylate serine-1 of troponin T.

scholarly journals 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.

1996 ◽  
Vol 16 (11) ◽  
pp. 6486-6493 ◽  
Author(s):  
Z Songyang ◽  
K P Lu ◽  
Y T Kwon ◽  
L H Tsai ◽  
O Filhol ◽  
...  
Keyword(s):  
Protein Kinases ◽  
Structural Basis ◽  
Phosphorylase Kinase ◽  
Primary Sequence ◽  
Casein Kinase I ◽  
Peptide Substrates ◽  
Substrate Specificities ◽  
Casein Kinases ◽  
Selection Of

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.

THE DYNACTOME: INVESTIGATING DYNAMIC KINASE NETWORKS IN CELLULAR DECISION-MAKING

2008 ◽  
Vol 31 (4) ◽  
pp. 22
Author(s):  
Jonathan So ◽  
Kelly Elder ◽  
Anna Dai ◽  
Claus Jorgensen ◽  
Rune Linding ◽  
...  
Keyword(s):  
Spectrometric Analysis ◽  
Phosphorylation Sites ◽  
Complex Samples ◽  
Downstream Signaling ◽  
Through Flow ◽  
Colorectal Cancer Cells ◽  
Induced Apoptosis ◽  
Cell Phenotypes ◽  
Kinase Phosphorylation ◽  
Kinase Expression

Networks of kinases play a role in the transmission and integration of signals from the membrane to the nucleus. We aim to elucidate kinase phosphorylation and interaction partners in these networks through the immuno-precipitation and mass spectrometric analysis of a representative set of 100 Flag-tagged kinases stably expressed in human colorectal cancer cells. The goal is to generate a comprehensive set of interactions and dynamic phosphorylation sites which correlate with cell phenotypes such as apoptosis and proliferation. The techniques of mass-spectrometry have allowed for the identification of proteins and their phosphorylation sites in complex samples. Various labeling methods such as iTRAQ has enabled the relative quantification of these sites as afunction of time (White et al. PNAS, 2007). However, kinases usually work in the context of particular signaling stimuli. We aim to characterize the role of these over-expressed kinases in the context of Trail-induced apoptosis. This isparticularly relevant to tumorigenesis in that many cancers are resistant to apoptosis and recombinant Trail therapies are currently undergoing clinical trials. We present assays to correlate the proliferative ability and sensitivity to apoptosis of various stable cell lines with kinase expression levels through flow cytometry. We also present efforts to trace downstream signaling through the monitoring of MAP kinase phosphorylation using a high-throughput bead array.

Sign in / Sign up

Export Citation Format

Share Document