Chemical Biology Approaches for Investigating the Functions of Lysine Acetyltransferases

Abstract

The side-chain acetylation of lysine residues in histones and non-histone proteins catalyzed by lysine acetyltransferases (KATs) represents a widespread posttranslational modification (PTM) in the eukaryotic cells. Lysine acetylation plays regulatory roles in major cellular pathways inside and outside the nucleus. In particular, KAT-mediated histone acetylation has an effect on all DNA-templated epigenetic processes. Aberrant expression and activation of KATs are commonly observed in human diseases, especially cancer. In recent years, the study of KAT functions in biology and disease has greatly benefited from chemical biology tools and strategies. In this Review, we present the past and current accomplishments in the design of chemical biology approaches for the interrogation of KAT activity and function. These methods and probes are classified according to their mechanisms of action and respective applications, with both strengths and limitations discussed.

Effect of lysine side chain length on histone lysine acetyltransferase catalysis

“…Given the challenges encountered in the development of KATs inhibitors, a better understanding of the substrate scope for KAT catalysis is of great biomolecular and medicinal interest. Exploring the substrate and cosubstrate specificity of KAT-catalyzed reactions is an important tool that can provide valuable information for the rational design of new inhibitors 31 . Towards this aim, extensive work has been carried out to unravel the specificity of the AcCoA cosubstrate in KAT catalysis 32,33 .…”

Identification of Lysine Isobutyrylation as A New Histone Modification Mark

“…We quantitatively measured the acylation activities of nine human KATs from three major KAT families, MYST, GCN5/PCAF/HAT1, and p300/CBP, using a fluorogenic assay that was designed to quantify the byproduct CoA. 27 Histone peptides H3 (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) or H4 (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) was used as the acyl acceptor to characterize the acetyl-, propionyl-, n-butyryl-, and isobutyrylation activities of individual KAT enzymes (Figure 3). Consistent with our recent study, 28 all the tested KAT enzymes show remarkable Kpr activity, more than 20% of their nascent acetyl transfer activity, which demonstrates that almost all eukaryotic KATs may possess this intrinsic activity.…”

“…[5][6][7][8] Lysine acetyltransferases (KATs) are the writer enzymes that introduce acylation marks on specific lysine residues using acyl-CoA molecules as the acyl donor. 9 Acyllysines are recognized by downstream reader proteins, and enzymatically reversed by eraser proteins, histone deacylases (HDAC). 10,11 Deregulation of lysine acylation dynamics caused by aberrant expression or mutation of either writer, reader, or eraser proteins is broadly associated with various disease phenotypes including inflammation, neurodegeneration, cancer, etc.…”

Bioorthogonal Reporters for Detecting and Profiling Protein Acetylation and Acylation

“…A dozen KAT members in mammalian cells have been identified and characterized both genetically and biochemically, which include the GNAT representative members GCN5, PCAF, and HAT1; five MYST family members (MOF, TIP60, MORF, MOZ, HBO1); p300 and CBP. [9][10][11][12][13][14][15][16] A few other proteins, such as CLOCK, NAT10, and NCOAT, though mentioned as KAT members, remain poorly characterized and are sometimes considered as orphan or noncanonical KAT members. 16,17 Importantly, many KATs were found to associate with cancer incidence, progression, and metastasis.…”

“…[9][10][11][12][13][14][15][16] A few other proteins, such as CLOCK, NAT10, and NCOAT, though mentioned as KAT members, remain poorly characterized and are sometimes considered as orphan or noncanonical KAT members. 16,17 Importantly, many KATs were found to associate with cancer incidence, progression, and metastasis. 15,18,19 Some KATs were found to efficiently catalyze other short-chain fatty acylation as well, especially protein propionylation and butyrylation.…”