Staphylococcus aureus produces hospital- and community-acquired infections, with methicillin-resistant S. posing a serious public health threat. The golden carotenoid pigment of aureus, staphyloxanthin, promotes resistance to reactive oxygen species host neutrophil-based killing, early enzymatic steps in staphyloxanthin production resemble those for cholesterol biosynthesis. We determined the crystal structures dehydrosqualene synthase (CrtM) at 1.58 angstrom resolution, finding structural...

Bisphosphonate drugs (e.g., Fosamax and Zometa) are thought to act primarily by inhibiting farnesyl diphosphate synthase (FPPS), resulting in decreased prenylation of small GTPases. Here, we show that some bisphosphonates can also inhibit geranylgeranyl (GGPPS), as well undecaprenyl (UPPS), a cis-prenyltransferase interest target for antibacterial therapy. Our results on GGPPS (10 structures) there three bisphosphonate-binding sites, consisting FPP or isopentenyl substrate-binding sites...

The gold color of Staphylococcus aureus is derived from the carotenoid staphyloxanthin, a virulence factor for organism. Here, we report synthesis and activity broad variety staphyloxanthin biosynthesis inhibitors that inhibit first committed step in its biosynthesis, condensation two farnesyl diphosphate (FPP) molecules to dehydrosqualene, catalyzed by enzyme dehydrosqualene synthase (CrtM). most active compounds are phosphonoacetamides have low nanomolar K(i) values CrtM inhibition whole...

β-Glucosidase (EC 3.2.1.21) plays an essential role in biofuel production since it can cleave β-1,4-glycosidic bond to convert cellobiose into fermentable glucose. Based on the structure of Trichoderma reesei β-glucosidase 2 (TrBgl2) we solved, amino acids outer channel active site were mutated this study. Mutants P172L and P172L/F250A showed most enhanced kcat/Km kcat values by 5.3- 6.9-fold, respectively, compared wild type (WT) toward 4-nitrophenyl-β-d-glucopyranoside (p-NPG) substrate at...

Laccases are multi-copper oxidases that catalyze the oxidation of various organic and inorganic compounds by reducing O2 to water. Here we report crystal structure at 1.8 Å resolution a native laccase (designated nLcc4) isolated from white-rot fungus Lentinus sp. nLcc4 is composed three cupredoxin-like domains D1-D3 each folded into Greek key β-barrel topology. T1 T2/T3 copper binding sites N-glycosylated Asn75, Asn238, Asn458 were elucidated. Initial rate kinetic analysis revealed kcat, Km,...

"Head-to-head" terpene synthases catalyze the first committed steps in sterol and carotenoid biosynthesis: condensation of two isoprenoid diphosphates to form cyclopropylcarbinyl diphosphates, followed by ring opening. Here, we report structures Staphylococcus aureus dehydrosqualene synthase (CrtM) complexed with its reaction intermediate, presqualene diphosphate (PSPP), (DHS) product, as well a series inhibitors. The results indicate that, on initial loss, primary carbocation so formed...

TcaR and IcaR are a weak strong negative regulator of transcription the ica locus, respectively, their presence prevents poly- N -acetylglucosamine production biofilm formation in Staphylococcus epidermidis . Although was shown to interact with promoter, precise binding region mechanism interaction remained unclear. Here we present 3D structure its apo form complex salicylate as well several aminoglycoside β-lactam antibiotics. A comparison native structures indicates that regulation...

Expression of the gene cluster icaADBC is necessary for biofilm production in Staphylococcus epidermidis . The ica operon negatively controlled by repressor IcaR. Here, crystal structure IcaR was determined and refined revealed a homodimer comprising entirely α-helices, typical tetracycline protein family regulations. N-terminal domain contains conserved helix-turn-helix DNA-binding motif with some conformational variations, indicating flexibility this region. C-terminal shows complementary...

A new type of glycan array covalently or noncovalently attached to aluminum oxide-coated glass (ACG) slides has been developed for studies enzymatic reactions and protein binding. To prepare the noncovalent array, glycans with a polyfluorinated hydrocarbon (−C8F17) tail are spotted robotically onto ACG slide surface containing layer terminated phosphonate. After incubation washing, can be characterized by MS-TOF via ionization/desorption at low laser energy without addition matrix....

Abstract Crown ethers are small, cyclic polyethers that have found wide‐spread use in phase‐transfer catalysis and, to a certain degree, protein chemistry. readily bind metallic and organic cations, including positively charged amino acid side chains. We elucidated the crystal structures of several protein‐crown ether co‐crystals grown presence 18‐crown‐6. then employed biophysical methods molecular dynamics simulations compare these complexes with corresponding apoproteins similar...

Squalene synthase (SQS) is a divalent metal-ion-dependent enzyme that catalyzes the two-step reductive `head-to-head' condensation of two molecules farnesyl pyrophosphate to form squalene using presqualene diphosphate (PSPP) as an intermediate. In this paper, structures human SQS and its mutants in complex with several substrate analogues intermediates coordinated Mg2+ or Mn2+ are presented, which stepwise delineate biosynthetic pathway. Extensive study active site has identified critical...

Abstract Epigenetic regulation is important for cancer progression; however, the underlying mechanisms, particularly those involving protein acetylation, remain to be fully understood. Here, we show that p300/ CBP ‐associated factor ( PCAF )‐dependent acetylation of transcription intestine‐specific homeobox ISX ) regulates epithelial–mesenchymal transition EMT and promotes metastasis. Mechanistically, at lysine 69 interaction with acetylated bromodomain‐containing 4 BRD 4) 332 in tumor...

Streptopain is a cysteine protease expressed by Streptococcus pyogenes. To study the maturation mechanism of streptopain, wild-type and Q186N, C192S, H340R, N356D W357A mutant proteins were in Escherichia coli purified to homogeneity. Proteolytic analyses showed that prostreptococcal pyrogenic exotoxin B zymogen (pro-SPE B) involves eight intermediates with combination ofcis- trans-processing. Based on sequences these intermediates, substrate specificity streptopain favors hydrophobic...

NkBgl, a β-glucosidase from Neotermes koshunensis, is β-retaining glycosyl hydrolase family 1 enzyme that cleaves β-glucosidic linkages in disaccharide or glucose-substituted molecules. β-Glucosidases have been widely used several applications. For example, mutagenesis of the attacking nucleophile has conducted to convert it into glycosynthase for synthesis oligosaccharides. Here, high-resolution structures wild-type mutated NkBgl complex with different ligand molecules are reported. In was...

ObjectivesAmino acid substitutions within the AdeRS two-component system are believed to result in overexpression of AdeABC efflux pump and extensive resistance antibiotics clinical Acinetobacter baumannii isolates. However, exact amino that cause remain unclear. We elucidated role by a complementation assay an adeRS knockout strain A. baumannii.

Autosomal dominant polycystic kidney disease (ADPKD) is the most common form of (PKD) and a typical adult-onset multisystem disorder. It progressive characterized by disruption renal tubular integrity, involving modulation cellular proliferation apoptosis. Most ADPKD results from mutation in either PKD1 or PKD2 gene encoding polycystin-1 polycystin-2, respectively. With inconsistent course ADPKD, biomarkers that can predict treatment efficacy rapid progression are needed. Studies have...

We have shown that Sp1 phosphorylation at Thr739 decreases its DNA-binding activity. In this study, we found of alone is necessary, but not sufficient for the inhibition activity during mitosis. demonstrated Pin1 could be recruited to Thr739(p)-Pro motif modulate interaction between phospho-Sp1 and CDK1, thereby facilitating CDK1-mediated Ser720, Thr723 Thr737 Loss C-terminal end (amino acids 741-785) significantly increased phosphorylation, implying C-terminus inhibits phosphorylation....

The extra-intestinal effects of probiotics for preventing allergic diseases are well known. However, the probiotic components that interact with host target molecules and have a beneficial effect on asthma remain unknown. Lactobacillus gasseri attenuates airway inflammation through activation peroxisome proliferator- activated receptor γ (PPARγ) in dendritic cells. Therefore, we aimed to isolate investigate immunomodulatory PPARγ component from L. gasseri.Culture supernatants were...

The MST family is a subclass of mammalian serine/threonine kinases that are related to the yeast sterile-20 protein and implicated in regulating cell growth transformation. MST3 contains 300-residue catalytic domain 130-residue regulatory domain, which can be cleaved by caspase activated autophosphorylation, promoting apoptosis. Here, five crystal structures presented, including complex with ADP manganese, unique cofactor preferred enzyme, adenine. Similar other kinases, folds into two...

Abstract Crown ethers are small, cyclic polyethers that have found wide‐spread use in phase‐transfer catalysis and, to a certain degree, protein chemistry. readily bind metallic and organic cations, including positively charged amino acid side chains. We elucidated the crystal structures of several protein‐crown ether co‐crystals grown presence 18‐crown‐6. then employed biophysical methods molecular dynamics simulations compare these complexes with corresponding apoproteins similar...