Molecular Characterization of Propionyllysines in Non-histone Proteins

Abstract

Lysine propionylation and butyrylation are protein modifications that were recently identified in histones. The molecular components involved in the two protein modification pathways are unknown, hindering further functional studies. Here we report identification of the first three in vivo non-histone protein substrates of lysine propionylation in eukaryotic cells: p53, p300, and CREB-binding protein. We used mass spectrometry to map lysine propionylation sites within these three proteins. We also identified the first two in vivo eukaryotic lysine propionyltransferases, p300 and CREB-binding protein, and the first eukaryotic depropionylase, Sirt1. p300 was able to perform autopropionylation on lysine residues in cells. Our results suggest that lysine propionylation, like lysine acetylation, is a dynamic and regulatory post-translational modification. Based on these observations, it appears that some enzymes are common to the lysine propionylation and lysine acetylation regulatory pathways. Our studies therefore identified first several important players in lysine propionylation pathway. Molecular & Cellular Proteomics 8:45-52, 2009.

Acylation of Biomolecules in Prokaryotes: a Widespread Strategy for the Control of Biological Function and Metabolic Stress

“…Although described as deacetylases, some sirtuins have depropionylase activity (56,57,102). In eukaryotes, certain sirtuins have also been shown to have protein desuccinylation (103,104) and demalonylation (105) activities and are thought to play a critical role in mitochondrial metabolism (reviewed in references 106 and 107).…”

“…For example, in Salmonella enterica, the activity of propionyl-CoA synthetase (PrpE) is controlled by propionylation (56). Both lysine propionylation and butyrylation have also been identified as reversible modifications that occur on histones (57,66), expanding the range of GNAT-mediated regulation through their ability to utilize various acyl-CoA substrates.…”

“…Their specificity is then due to their structurally divergent N-and C-terminal domains outside the core domain (49). In some cases, other acyl-CoA thioesters, such as propionyl-CoA and succinyl-CoA, are physiologically relevant substrates (43,56,57) and are discussed in more detail below.…”

Succinylome Analysis Reveals the Involvement of Lysine Succinylation in Metabolism in Pathogenic Mycobacterium tuberculosis*

“…Among the hundreds of different PTMs, acylations at lysine residues, such as acetylation (3)(4)(5)(6), malonylation (7,8), crotonylation (9,10), propionylation (11)(12)(13), butyrylation (11,13), and succinylation (7, 14 -16) are crucial for functional regulations of many prokaryotic and eukaryotic proteins. Because these lysine PTMs depend on the acyl-CoA metabolic intermediates, such as acetyl-CoA (Ac-CoA), succinyl-CoA, and malonylCoA, lysine acylation could provide a mechanism to respond to changes in the energy status of the cell and regulate energy metabolism and the key metabolic pathways in diverse organisms (17,18).…”

Lysine Malonylation Is Elevated in Type 2 Diabetic Mouse Models and Enriched in Metabolic Associated Proteins

“…For example, adenosine-triphosphate (ATP) is used in phosphorylation, S-adenosylmethionine (SAM) in methylation, and acetyl-CoA in acetylation. Lysine acylations including malonylation (6), succinylation (7), butyrylation (8), propionylation (9), and crotonylation (10) represent a group of PTMs that use intermediates of energy metabolism like malonyl-CoA, succinyl-CoA, butyryl-CoA, propionyl-CoA, and crotonyl-CoA as group donors. Among the lysine acylations, lysine malonylation was first identified in Escherichia coli (E. coli) and HeLa cells using a specific anti-Kmal (anti-malonyllysine) antibody (6).…”