Deciphering Enzyme Function Using Peptide Arrays

Abstract

Enzymes are key molecules in signal-transduction pathways. However, only a small fraction of more than 500 human kinases, 300 human proteases and 200 human phosphatases is characterised so far. Peptide microarray based technologies for extremely efficient profiling of enzyme substrate specificity emerged in the last years. This technology reduces set-up time for HTS assays and allows the identification of downstream targets. Moreover, peptide microarrays enable optimisation of enzyme substrates. Focus of this review is on assay principles for measuring activities of kinases, phosphatases or proteases and on substrate identification/optimisation for kinases. Additionally, several examples for reliable identification of substrates for lysine methyl-transferases, histone deacetylases and SUMO-transferases are given. Finally, use of high-density peptide microarrays for the simultaneous profiling of kinase activities in complex biological samples like cell lysates or lysates of complete organisms is described. All published examples of peptide arrays used for enzyme profiling are summarised comprehensively.

Tumor phosphatidylinositol 3-kinase signaling in therapy resistance and metastatic dissemination of rectal cancer: Opportunities for signaling-adapted therapies

“…Ideally, to become clinically useful, technologies must provide rapid and high-fidelity analysis of small-quantity samples. Kinase substrate array technologies are high-throughput tools for global profiling of kinase activities in tissue samples without prior knowledge of which signaling pathways are activated [33][34][35][36]. Using such arrays, each tumor sample will generate an individual phosphosubstrate signature, theoretically portraying the state of composite information flow through signaling cascades.…”

Peptide Microarrays for Profiling of Epigenetic Targets

“…Generally, there are two strategies, differing in the nature of the cosubstrate, used for the read-out of enzymatic activities on peptide microarrays comprehensively reviewed in [66,67] (Figure 9.4). Generally, there are two strategies, differing in the nature of the cosubstrate, used for the read-out of enzymatic activities on peptide microarrays comprehensively reviewed in [66,67] (Figure 9.4).…”

Combining SAMDI Mass Spectrometry and Peptide Arrays to Profile Phosphatase Activities

“…Peptide arrays have emerged as an approach for studying the substrate specificities of a variety of enzymes (Arsenault, Griebel, & Napper, 2011; Foong, Fu, Yao, & Uttamchandani, 2012; Katz et al, 2011; Szymczak, Kuo, & Mrksich, 2017; Thiele, Stangl, & Schutkowski, 2011). Peptide arrays allow enzyme activities to be measured on a broad range of substrates, giving a more complete understanding of specificity.…”