Summary Acyl‐ CoA and acyl‐acyl carrier protein ( ACP ) synthetases activate exogenous fatty acids for incorporation into phospholipids in G ram‐negative bacteria. However, Gram‐positive bacteria utilize an acyltransferase pathway the biogenesis of phosphatidic acid that begins with acylation sn ‐glycerol‐3‐phosphate by PlsY using acyl‐phosphate (acyl‐ PO 4 intermediate. PlsX generates acyl‐ from end‐products synthesis. The plsX gene S taphylococcus aureus was inactivated resulting strain...

Pantothenate kinase (PANK) is a metabolic enzyme that regulates cellular coenzyme A (CoA) levels. There are three human PANK genes, and inactivating mutations in PANK2 lead to pantothenate associated neurodegeneration (PKAN). Here we performed library screen followed by chemical optimization produce PZ-2891, an allosteric activator crosses the blood brain barrier. PZ-2891 occupies pocket engages dimer interface form PANK•ATP•Mg2+•PZ-2891 complex. The binding of one protomer locks opposite...

Abstract The interaction between SStp, the transit peptide of precursor protein to small subunit Rubisco (prSSU) and two Hsp70 molecular chaperones, Escherichia coli DnaK pea (Pisum sativum) CSS1, was investigated in detail. Two statistical analyses were developed used investigate predict regions SStp recognized by DnaK. Both algorithms suggested that would have high affinity for N terminus moderate central region, low C terminus. Furthermore, both predicted this pattern >75% peptides...

Summary Bacteria respond to physical and chemical stresses that affect the integrity of cell wall membrane by activating an intricate envelope stress response. The ability cells regulate biophysical properties adjusting fatty acid composition is known as homeoviscous adaptation. Here, we identify a adaptation mechanism in Bacillus subtilis regulated extracytoplasmic function σ factor W . Cell active compounds, including detergents, activate sense‐oriented, ‐dependent promoter within first...

Oleate hydratases (OhyAs) belong to a large family of bacterial proteins catalyzing the hydration or isomerization double bonds in unsaturated fatty acids. A Staphylococcus aureus gene (Sa0102) is predicted encode an OhyA. Here, we recombinantly expressed and purified SaOhyA found that it forms homodimer requires FAD for activity. hydrates only acids containing cis-9 bonds, but not with trans-9 cis at other positions. products were detected S. phospholipids released into growth medium. does...

utilizes the fatty acid (FA) kinase system to activate exogenous FAs for membrane synthesis. We developed a lipidomics workflow determine phosphatidylglycerol (PG) molecular species synthesized by

Abstract Threonine deaminase (IlvA) is the first enzyme in isoleucine branch of branched-chain amino acid biosynthetic pathway. Kinetic studies feedback inhibition Escherichia coli IlvA (EcIlvA) introduced concept allosteric regulation enzymes. While crystal structure EcIlvA shows a tetrameric assembly protomers with distinct catalytic and regulatory domains, binding site within domain (EcIlvA_R) its mechanism remained undefined. Here, we present high-resolution EcIlvA_R•Ile complex,...

Gram-positive bacteria, including Listeria monocytogenes, adjust membrane fluidity by shortening the fatty acid chain length and increasing proportional production of anteiso acids at lower growth temperatures. The first condensation reaction in biosynthesis is carried out beta-ketoacyl-acyl carrier protein synthase III (FabH), which determines type produced bacteria. Here, we measured initial rates FabH-catalyzed malonyl-acyl alternate branched-chain precursor acyl-CoAs utilizing...

The SaeRS two-component system is a master activator of virulence factor transcription in

Pantothenate kinase is the master regulator of CoA biosynthesis and feedback-inhibited by acetyl-CoA. Comparison human PANK3·acetyl-CoA complex to structures PANK3 in four catalytically relevant complexes, 5′-adenylyl-β,γ-imidodiphosphate (AMPPNP)·Mg2+, AMPPNP·Mg2+·pantothenate, ADP·Mg2+·phosphopantothenate, AMP phosphoramidate (AMPPN)·Mg2+, revealed a large conformational change dimeric enzyme. The amino-terminal nucleotide binding domain rotates close active site, this allows P-loop engage...

Fatty acid (FA) kinase produces acyl-phosphate for the synthesis of membrane phospholipids in Gram-positive bacterial pathogens. FA consists a protein (FakA) that phosphorylates an substrate bound to second module, FA-binding (FakB). Staphylococcus aureus expresses two distinct, but related, FakBs with different selectivities. Here, we report structures FakB1 four saturated FAs at 1.6-1.93 Å resolution. We observed are accommodated within slightly curved hydrophobic cavity whose length is...

Therapeutic relief of CoA sequestration improves mitochondrial dysfunction in a propionic acidemia mouse model.

Lysophospholipids are deacylated derivatives of their bilayer forming phospholipid counterparts that present at low concentrations in cells. Phosphatidylglycerol (PG) is the principal membrane Staphylococcus aureus and lysophosphatidylglycerol (LPG) detected abundance. Here, we used a mass spectrometry screen to identify locus SAUSA300_1020 as gene responsible for maintaining 1-acyl-LPG S. aureus. The encodes protein with predicted amino terminal transmembrane α-helix attached globular...

Pantothenate kinase (PANK) is the critical regulator of intracellular levels coenzyme A and has emerged as an attractive target for treating neurological metabolic disorders. This report describes optimization, synthesis, full structure-activity relationships a new chemical series pantothenate competitive PANK inhibitors. Potent drug-like molecules were obtained by optimizing high throughput screening hit, using lipophilic ligand efficiency (LipE) derived from human PANK3 IC50 values to...

Emerging research has revealed that the cellular coenzyme A (CoA) supply can become limiting with a detrimental impact on growth, metabolism and survival. Measurement of CoA is challenge due to its relatively low abundance dynamic conversion free thioesters that, in turn, participate numerous metabolic reactions. method described navigates through potential pitfalls during sample preparation yield an assay broad linear range detection suitable for use many biomedical laboratories.

A Schiff base ligand L was synthesized by condensation of 1,2-diaminoethane with creatinine. The reaction the metal chloride salt gives Co(II) and Cu(II) complexes. its complexes were characterized elemental analysis, FT-IR, NMR, UV?Vis, conductivity magnetic susceptibility measurements as well thermal analyses. Based on spectral data, tetrahedral geometries have been proposed for molar data show that are non-electrolytic in nature. In DFT studies, geometry fully optimized using B3LYP...

Conversion of pantothenate to phosphopantothenate in humans is the first dedicated step coenzyme A (CoA) biosynthesis pathway and mediated by four isoforms kinase. These enzymes are allosterically regulated acyl-CoA levels, which control rate CoA biosynthesis. Small molecule activators PANK that overcome feedback suppression increase levels cultured cells animals have shown great potential for treatment kinase-associated neurodegeneration propionic acidemias. In this study, we detail further...

Abstract Histone acetyltransferases (HAT) catalyze the acylation of lysine side chains and are implicated in diverse human cancers as both oncogenes non-oncogene dependencies 1 . Acetyl-CoA-competitive HAT inhibitors have garnered attention potential cancer therapeutics first clinical trial for this class is ongoing ( NCT04606446 ). Despite broad enthusiasm these targets, notably including CBP/p300 KAT6A/B 2–5 , mechanisms therapeutic response evolved drug resistance remain poorly...

The central position of coenzyme A (CoA) in intermediary metabolism and its role human neurodegenerative disease, led us to develop chemical probes targeting pantothenate kinase (PANK), the regulatory enzyme that controls CoA biosynthesis. There are four mammalian PANK isoforms exhibit tissue‐specific expression differ their sensitivity feedback inhibition by thioesters. high‐throughput screen further medicinal chemistry efforts identified a novel class orally bioavailable PanK modulators...