In Vivo and in Vitro Analyses of Single-amino Acid Variants of the Salmonella enterica Phosphotransacetylase Enzyme Provide Insights into the Function of Its N-terminal Domain

Brinsmade, Shaun R.; Escalante‐Semerena, Jorge C.

doi:10.1074/jbc.m611439200

Cited by 17 publications

(41 citation statements)

References 66 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cells were harvested and MBP-H 6 -PncA purified as described for purification of GST-H 6 -Pat. PncA eluted at ϳ30% buffer B. MBP-H 6 -PncA-containing fractions were pooled and H 6 -rTEV protease (26) added to reach a 1:50 H 6 -rTEV protease:MBP-H 6 -PncA ratio; H 6 -rTEV protease was purified as described (27). The cleavage reaction mixture was incubated at room temperature for 3 h and dialyzed overnight against two liters of buffer A at 4°C.…”

Section: Protein Purificationmentioning

confidence: 99%

N-Lysine Propionylation Controls the Activity of Propionyl-CoA Synthetase

Garrity

Gardner

Hawse

et al. 2007

Journal of Biological Chemistry

Self Cite

188

229

View full text Add to dashboard Cite

Reversible protein acetylation is a ubiquitous means for the rapid control of diverse cellular processes. Acetyltransferase enzymes transfer the acetyl group from acetyl-CoA to lysine residues, while deacetylase enzymes catalyze removal of the acetyl group by hydrolysis or by an NAD ؉ -dependent reaction. Propionyl-coenzyme A (CoA), like acetyl-CoA, is a high energy product of fatty acid metabolism and is produced through a similar chemical reaction. Because acetyl-CoA is the donor molecule for protein acetylation, we investigated whether proteins can be propionylated in vivo, using propionyl-CoA as the donor molecule. We report that the Salmonella enterica propionyl-CoA synthetase enzyme PrpE is propionylated in vivo at lysine 592; propionylation inactivates PrpE. The propionyl-lysine modification is introduced by bacterial Gcn-5-related N-acetyltransferase enzymes and can be removed by bacterial and human Sir2 enzymes (sirtuins). Like the sirtuin deacetylation reaction, sirtuin-catalyzed depropionylation is NAD ؉ -dependent and produces a byproduct, O-propionyl ADP-ribose, analogous to the O-acetyl ADP-ribose sirtuin product of deacetylation. Only a subset of the human sirtuins with deacetylase activity could also depropionylate substrate. The regulation of cellular propionylCoA by propionylation of PrpE parallels regulation of acetylCoA by acetylation of acetyl-CoA synthetase and raises the possibility that propionylation may serve as a regulatory modification in higher organisms.Protein acetylation is a ubiquitous means for the rapid control of diverse cellular processes (1, 2). Acetylation occurs at lysine residues, with acetyl-CoA (Ac-CoA) 5 serving as the acetyl group donor. In higher organisms, aberrant acetylation of lysine residues in histone tails correlates with diseases such as cancers and developmental disorders and may contribute to modulation of cell life span (3, 4). From bacteria to humans, N-Lys acetylation controls the activity of acetyl-coenzyme A synthetase (AMP-forming; Acs) and potentially that of other members of the AMP-forming family of enzymes (5-7). In Salmonella enterica, Acs is deacetylated by CobB, a member of the Sir2 family of NAD ϩ -dependent deacetylases (a.k.a. sirtuins) (8). Interestingly, strains of S. enterica lacking CobB deacetylase activity cannot grow on propionate because the propionyl-CoA synthetase (encoded by the prpE gene) that activates propionate to propionyl-CoA is not active (5, 9).Cells generate propionyl-CoA from several different processes, including the catabolism of odd chain fatty acids, the decarboxylation of succinate, the catabolism of amino acids (e.g. threonine), and the activation of propionate (10 -12). Propionate is a powerful inhibitor of cell growth that is widely used as a food preservative. Reports in the literature suggest that propionyl-CoA may be responsible for the cytotoxic effects of propionate. Although the direct effects of propionyl-CoA are unclear, it is clear that cells avoid accumulating this metabolite (13-15). The cell maintains...

show abstract

Section: Protein Purificationmentioning

confidence: 99%

N-Lysine Propionylation Controls the Activity of Propionyl-CoA Synthetase

Garrity

Gardner

Hawse

et al. 2007

Journal of Biological Chemistry

Self Cite

188

229

View full text Add to dashboard Cite

show abstract

“…So far, only two PtaIIs from E. coli and S. enterica have been shown to be allosterically regulated via their N-terminal domains by NADH, ATP and pyruvate [5,6]. In this study, we also first time showed that a-ketoglutarate, a substrate of a-ketoglutarate dehydrogenase complex (a-KGDC), one of the rate determining enzymes of TCA cycle, allosterically inhibited recombinant S. saprophyticus Pta activity in both forward and reverse directions.…”

Section: Allosteric Inhibition Of S Saprophyticus Pta Activity By α-mentioning

confidence: 57%

“…PtaII, on the other hand, is encoded by the pta gene, which is organized in a single operon along with the ackA gene. PtaIIs contain three conserved domains: the P loop NTPase domain at the N-terminus, a DRTGG domain and the C-terminal domain where active site is located [5,6]. The C-terminal domain shares high homology with PtaIs.…”

Section: Introductionmentioning

confidence: 99%

“…The C-terminal domain shares high homology with PtaIs. The P loop NTPase domain, on the other hand, acts as a regulatory domain whereby the allosteric effectors, namely NADH, ATP and pyruvate, display their effects on the activity [5,6]. Unlike PtaIIs, PtaIs have been considered not to be allosterically regulated.…”

Section: Introductionmentioning

confidence: 99%

“…So far, two classes of Ptas have been identified: PtaIs and PtaIIs, which are about 350 and 700 residues in length, respectively [3][4][5]. PtaI is encoded by the eutD gene 1 .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A phosphotransacetylase from uropathogenic Staphylococcus saprophyticus: Characterization, kinetic analysis and its allosteric inhibition by α-ketoglutarate

Aksoy¹

2012

Turk J Bioch

View full text Add to dashboard Cite

Microbial production of odd‐chain fatty acids

Qin

Wang

et al. 2022

Biotech & Bioengineering

View full text Add to dashboard Cite

Odd-chain fatty acids (OcFAs) and their derivatives have attracted much attention due to their beneficial physiological effects and their potential to be alternatives to advanced fuels. However, cells naturally produce even-chain fatty acids (EcFAs) with negligible OcFAs. In the process of biosynthesis of fatty acids (FAs), the acetyl-CoA serves as the starter unit for EcFAs, and propionyl-CoA works as the starter unit for OcFAs. The lack of sufficient propionyl-CoA, the precursor, is usually regarded as the main restriction for large-scale bioproduction of OcFAs. In recent years, synthetic biology strategies have been used to modify several microorganisms to produce more propionyl-CoA that would enable an efficient biosynthesis of OcFAs. This review discusses several reported and potential metabolic pathways for propionyl-CoA biosynthesis, followed by advances in engineering several cell factories for OcFAs production. Finally, trends and challenges of synthetic biology driven OcFAs production are discussed.

show abstract

In Vivo and in Vitro Analyses of Single-amino Acid Variants of the Salmonella enterica Phosphotransacetylase Enzyme Provide Insights into the Function of Its N-terminal Domain

Cited by 17 publications

References 66 publications

N-Lysine Propionylation Controls the Activity of Propionyl-CoA Synthetase

N-Lysine Propionylation Controls the Activity of Propionyl-CoA Synthetase

A phosphotransacetylase from uropathogenic Staphylococcus saprophyticus: Characterization, kinetic analysis and its allosteric inhibition by α-ketoglutarate

Microbial production of odd‐chain fatty acids

Contact Info

Product

Resources

About