+ Site Statistics
References:
54,258,434
Abstracts:
29,560,870
PMIDs:
28,072,757
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

Calcium-dependent protein kinases from Arabidopsis show substrate specificity differences in an analysis of 103 substrates



Calcium-dependent protein kinases from Arabidopsis show substrate specificity differences in an analysis of 103 substrates



Frontiers in Plant Science 2: 36



The identification of substrates represents a critical challenge for understanding any protein kinase-based signal transduction pathway. In Arabidopsis, there are more than 1000 different protein kinases, 34 of which belong to a family of Ca(2+)-dependent protein kinases (CPKs). While CPKs are implicated in regulating diverse aspects of plant biology, from ion transport to transcription, relatively little is known about isoform-specific differences in substrate specificity, or the number of phosphorylation targets. Here, in vitro kinase assays were used to compare phosphorylation targets of four CPKs from Arabidopsis (CPK1, 10, 16, and 34). Significant differences in substrate specificity for each kinase were revealed by assays using 103 different substrates. For example CPK16 phosphorylated Serine 109 in a peptide from the stress-regulated protein, Di19-2 with K(M) ∼70 μM, but this site was not phosphorylated significantly by CPKs 1, 10, or 34. In contrast, CPKs 1, 10, and 34 phosphorylated 93 other peptide substrates not recognized by CPK16. Examples of substrate specificity differences among all four CPKs were verified by kinetic analyses. To test the correlation between in vivo phosphorylation events and in vitro kinase activities, assays were performed with 274 synthetic peptides that contained phosphorylation sites previously mapped in proteins isolated from plants (in vivo-mapped sites). Of these, 74 (27%) were found to be phosphorylated by at least one of the four CPKs tested. This 27% success rate validates a robust strategy for linking the activities of specific kinases, such as CPKs, to the thousands of in planta phosphorylation sites that are being uncovered by emerging technologies.

(PDF emailed within 0-6 h: $19.90)

Accession: 051907981

Download citation: RISBibTeXText

PMID: 22645532

DOI: 10.3389/fpls.2011.00036


Related references

Differential centrifugation analysis of subcellular distribution of substrates of cyclic amp dependent calcium calmodulin dependent and calcium phospholipid dependent protein kinases in rat sertoli cells. Biology of Reproduction 36(SUPPL 1): 143, 1987

Substrate specificity of calcium cam dependent multifunctional protein kinases comparison of isoenzymes from brain liver and skeletal muscle. Biochemical & Biophysical Research Communications 151(3): 1332-1338, 1988

Substrate specificity of protein kinases and computational prediction of substrates. Biochimica et Biophysica Acta 1754(1-2): 200-209, 2005

Mitogen-activated protein kinases and calcium-dependent protein kinases are involved in wounding-induced ethylene biosynthesis in Arabidopsis thaliana. Plant, Cell and Environment 41(1): 134-147, 2017

Endogenous substrate proteins for calcium calmodulin dependent calcium phospho lipid dependent and cyclic amp dependent protein kinases in mouse pancreatic islets. Biochemical Journal 221(1): 247-254, 1984

Existence of endogenous substrate proteins for calcium calmodulin dependent and calcium phospholipid dependent protein kinases in rat adrenal glomerulosa cells. Journal of Steroid Biochemistry 29(3): 277-284, 1988

The substrate specificity of the protein kinase induced in cells infected with herpesviruses: studies with synthetic substrates [corrected] indicate structural requirements distinct from other protein kinases. Biochimica et Biophysica Acta 889(2): 208-215, 1986

Calcium signaling through protein kinases. The Arabidopsis calcium-dependent protein kinase gene family. Plant Physiology 129(2): 469-485, 2002

Characterization of Arabidopsis calcium-dependent protein kinases: activated or not by calcium?. Biochemical Journal 447(2): 291-299, 2012

Sequence characteristics, subcellular localization, and substrate specificity of DYRK-related kinases, a novel family of dual specificity protein kinases. Journal of Biological Chemistry 273(40): 25893-25902, 1998

Differential substrate specificity of a calcium phosphatidylserine diglyceride dependent and a novel calcium independent ps dg dependent protein kinase in human neutrophils. Clinical Research 38(2): 434A, 1990

Myristoylation of calcium-dependent protein kinases in Arabidopsis. Plant Biology (Rockville) : 57, 1999

Membrane association of calcium-dependent protein kinases in Arabidopsis. Plant Biology (Rockville) : 36, 2001

Myristoylation of calcium-dependent protein kinases from Arabidopsis thaliana. 2005

Plasma membrane localized calcium-dependent protein kinases in Arabidopsis. Plant Biology (Rockville) : 177, 2002