The Acyl-CoA Specificity of Human Lysine Acetyltransferase KAT2A

Abstract

Protein post-translational modifications serve to regulate a broad range of cellular functions including signal transduction, transcription, and metabolism. Protein lysine residues undergo many post-translational acylations and are regulated by a range of enzymes, such as histone acetyl transferases (HATs) and histone deacetylases (HDACs). KAT2A, well characterized as a lysine acetyltransferase for both histone and nonhistone substrates, has been reported to tolerate additional acyl-CoA substrates, such as succinyl-CoA, and shows nonacetyl transferase activity in specific biological contexts. In this work, we investigate the acyl-CoA substrate preference of KAT2A and attempt to determine whether and to what extent additional acyl-CoA substrates may be utilized by KAT2A in a cellular context. We show that while KAT2A can bind and utilize malonyl-CoA, its activity with succinyl-CoA or glutaryl-CoA is very weak, and acetylation is still the most efficient activity for KAT2A in vitro and in cells.

Metabolomic, proteomic and single cell proteomic analysis of cancer cells treated with the KRASG12Dinhibitor MRTX1133

“…These results suggest that the analysis of histone PTMs in single human cells would be a practical avenue for research. However, as in traditional proteomics, the use of alternative proteases 24 or lysine derivatization prior to tryptic digestion 25 would be required for accurate assignment of the PTM to the originating proteoform.…”

Efficient Chemical and Enzymatic Syntheses of FAD Nucleobase Analogues and Their Analysis as Enzyme Cofactors**

“…The chemical and enzymatic synthesis methods of FAD nucleobase analogues we report here form the basis of production of artificial fluorescent and other non‐natural FAD variants in the future. These FAD analogues, similar to SAM and NAD + analogues reported in literature, may have interesting applications in drug development, cell imaging, and biotechnology applications such as bioremediation [43–51] . For example, a diverse library of SAM analogues was created and tested as cofactors in methyltransferase enzymes implicated in small molecule and RNA methylations [51] .…”

“…These FAD analogues, similar to SAM and NAD + analogues reported in literature, may have interesting applications in drug development, cell imaging, and biotechnology applications such as bioremediation. [43][44][45][46][47][48][49][50][51] For example, a diverse library of SAM analogues was created and tested as cofactors in methyltransferase enzymes implicated in small molecule and RNA methylations. [51] In another instance, cytosine analogues of NAD + have been used as biorthogonal redox systems with engineered enzymes formate dehydrogenase and malic enzyme for CO 2 fixation in E. coli.…”

New insights into the pathophysiology of methylmalonic acidemia

“…In addition to traditional genetic and biological techniques, going forward we envision chemical biology could play a critical role in defining and manipulating aberrant acylation in MMA and related disorders. Unlike canonical enzyme‐catalyzed histone acetylation, substantial evidence indicates that SIRT5‐regulated protein malonylation, succinylation, and glutarylation is driven nonenzymatically by the reactivity of their acyl‐CoA precursors 65,107–110 . In particular, the high reactivity of malonyl‐CoA has been attributed to the electron‐withdrawing effect of the β‐carboxy group, an attribute that should also extend to methylmalonyl‐CoA 111,112 .…”

“…Unlike canonical enzyme-catalyzed histone acetylation, substantial evidence indicates that SIRT5-regulated protein malonylation, succinylation, and glutarylation is driven nonenzymatically by the reactivity of their acyl-CoA precursors. 65,[107][108][109][110] In particular, the high reactivity of malonyl-CoA has been attributed to the electron-withdrawing effect of the β-carboxy group, an attribute that should also extend to methylmalonyl-CoA. 111,112 In the future we envision that chemoproteomic studies of thioester reactivity may be helpful in identifying new protein targets of aberrant hyperacylation.…”