An atlas of the catalytically active liver and spleen kinases in chicken identified by chemoproteomics

Bindu Nanduri; Cathy R Gresham; Winnie W Hui; Mark Ou; Richard H Bailey; Mariola J Edelmann

doi:10.1016/j.jprot.2020.103850

An atlas of the catalytically active liver and spleen kinases in chicken identified by chemoproteomics

J Proteomics. 2020 Aug 15:225:103850. doi: 10.1016/j.jprot.2020.103850. Epub 2020 Jun 2.

Authors

Bindu Nanduri¹, Cathy R Gresham², Winnie W Hui³, Mark Ou³, Richard H Bailey⁴, Mariola J Edelmann⁵

Affiliations

¹ Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS, USA; Institute for Genomics, Biocomputing, and Biotechnology, Mississippi State University, Mississippi State, MS, USA.
² Institute for Genomics, Biocomputing, and Biotechnology, Mississippi State University, Mississippi State, MS, USA.
³ Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, USA.
⁴ Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS, USA.
⁵ Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, USA. Electronic address: medelmann@ufl.edu.

PMID: 32502695
DOI: 10.1016/j.jprot.2020.103850

Abstract

Phosphorylation is a post-translational protein modification regulating most known cellular processes. While protein kinases constitute a large family of highly conserved enzymes, identification of active kinases is challenging due to a low abundance of some of these signaling molecules. Although chicken is the first agricultural animal to have a sequenced genome, annotation of the kinome, i.e., a complement of all protein kinases in the genome is limited. We used chemical probes consisting of ATP and ADP derivatives binding to specific lysine (Lys) residues within the ATP-binding pocket of kinases, combined with proteomics, to identify 267 peptides labeled with the ATP and ADP acyl derivatives and 188 corresponding chicken kinases in chicken spleen and liver. Our description of active chicken kinases and ATP binding sites will support future studies focused on identifying the role of this important class of enzymes in chicken health and disease. SIGNIFICANCE: Advances made in understanding chicken enzymes are critical for the improved knowledge of the regulatory pathways controlling physiological processes in chicken. Since protein phosphorylation controls multiple aspects of cell fate, it is often linked to pathological conditions, and understanding of the kinase expression in chicken is essential for future therapeutic approaches. We coupled proteomics and labeling with active-site probes binding to Lys residues within the ATP-binding pocket of kinases to identify 188 kinases and corresponding 267 peptides labeled with the ATP and ADP acyl derivatives in chicken spleen and liver. Results of the present study describing catalytically active kinases is a starting point for chemoproteomic-based interrogation of kinases in chicken exposed to different conditions. Kinases identified in this study are available through the Chickspress genome browser that has previously published mRNA, miRNA, and shotgun proteomics data.

Keywords: Chemoproteomics; Chicken; Gallus gallus; Kinome; Protein kinase.

MeSH terms

Adenosine Triphosphate
Animals
Chickens*
Liver
Protein Kinases
Proteomics
Spleen*

Substances

Adenosine Triphosphate
Protein Kinases