Abstract
Recently, peptide microarrays have been used to distinguish proteins, antibodies, viruses, and bacteria based on their binding to random sequence peptides. We reported on the use of peptide arrays to identify enzyme modulators that involve screening an array of 10,000 defined and addressable peptides on a microarray. Primary peptides were first selected to inhibit the enzyme at low μM concentrations. Then, new peptides were found to only bind strongly with the enzyme–inhibitor complex, but not the native enzyme. These new peptides served as secondary inhibitors that enhanced the inhibition of the enzyme together with the primary peptides. Without the primary peptides, the secondary effect peptides had little effect on the enzyme activity. Conversely, we also selected peptides that recovered the activities of inhibited enzyme–peptide complex. The selection of cooperative peptide pairs will provide a versatile toolkit for modulating enzyme functions, which may potentially be applied to drug discovery and biocatalysis.
Highlights
Small molecules that regulate enzyme activity play an important role in many biological functions, and are crucial for drug discovery [1,2]
To stabilize PEP-1–β-Gal complex with inhibited enzyme activity, we used formaldehyde to covalently crosslink the peptide with the enzyme surface that permanently inhibited enzymes
The primary peptide inhibitor was crosslinked with enzymes to stabilize the inhibited peptide–enzyme complex
Summary
Small molecules that regulate enzyme activity play an important role in many biological functions, and are crucial for drug discovery [1,2]. Peptides represent a promising class of potential enzyme modulators [5] due to their large chemical diversity [6] and the existence of well-established approaches for library synthesis [7]. Peptides and their derivatives are found to inhibit many important enzymes [8], such as dehydrogenases [9], protein kinases [10], and proteases [11]. Over the past few decades, peptide microarrays have been developed for the high-throughput screening of a library of peptides that can bind to biological targets and alter their functions [7].
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