Phosphoproteome and Kinome Analysis: Unique Perspectives on the Same Problem

Abstract

Since ancient times many natural organic materials have been used in artistic and historic works as binders, adhesives, fillers and coatings. The identification of these materials is important not only for a proof of authenticity; for restorers and conservators it is essential to recognize the materials and technologies employed by artists and craftsmen. For identification and characterisation of the different natural organic binders spectrometric and chromatographic methods are well established. Recently, capillary electrophoresis has been introduced as an alternative technique to receive analytical information about the kind and composition of the binding media used in artefacts. These materials are waxes, resins, drying oils, animal glues and plant gums. In the present review the application of capillary electrophoresis in this area is discussed against the background of a general survey of the more common analytical techniques.

Genome to Kinome: Species-Specific Peptide Arrays for Kinome Analysis

“…Blastp results revealed that about half of the bovine sequences had 100% identity to the query sequences and that the majority of the remaining peptides had limited sequence divergence (Table 1). A comparison of the protein descriptions for the query and hit sequences confirmed that they referred to orthologous proteins (19). These results indicate that phosphorylation sites between human and bovine orthologous proteins are not absolutely conserved, which suggests that a human array would be of limited value for bovine kinome analysis.…”

Systems Kinomics Demonstrates Congo Basin Monkeypox Virus Infection Selectively Modulates Host Cell Signaling Responses as Compared to West African Monkeypox Virus

“…Thus, we employed systems kinomics (20,29,30) with high-throughput human kinome peptide arrays to study the global activation state of host kinases, the kinome, following West African or Congo Basin MPXV infection. The resultant kinome data sets were subjected to hierarchical cluster analysis for comparative analysis and visualization of changes to host cell phosphorylation following infection.…”

Peptide arrays for kinome analysis: New opportunities and remaining challenges

“…A number of experimental approaches are available for the analysis of phosphorylation based cellular signaling, these can be divided into two groups, phosphoproteome and kinome analysis 16. The difference is dependent on whether the consideration is on the protein kinases that phosphorylate proteins, the kinome, or the targets of these enzymes, the phosphoproteome.…”

“…This is exacerbated by the fact that these proteins, in addition to being found in low abundance, are often phosphorylated at sub‐stoichiometric levels. This means that a small fraction of a given signaling protein is phosphorylated at any one time; only 1–2% of the total individual protein compliment of a cell is found in the phosphorylated form 16–19. When considering all these experimental limitations, a promising alternative for characterization of cellular phosphorylation is to focus instead on the kinome.…”

“…The basic premise of kinome analysis through peptide arrays is that peptides representing the phosphorylation target sites of proteins are synthesized and spotted onto an array surface 23. Detailed review of the commonly employed methodologies for peptide synthesis and array spotting are available elsewhere 16. Following production of the array a sample containing active kinase, or kinases, such as a cellular lysate, is applied for the array and these enzymes phosphorylate their respective target sequences using ATP as the phosphate source.…”