Comparison of apple polyphenol-gelatin binary complex and apple polyphenol-gelatin-pectin ternary complex: Antioxidant and structural characterization

Abstract The large subunit of phage lambda terminase, gpA, the gene product of the phage A gene, interacts with the small subunit, gpNul, to form functional terminase. Terminase binds to lambda DNA at cosB to form a binary complex. The terminase:DNA complex binds a prohead to form a ternary complex. Ternary complex formation involves an interaction of the prohead with gpA. The amino terminus of gpA contains a functional domain for interaction with gpNul, and the carboxy-terminal 38 amino acids of gpA contain a functional domain for prohead binding. This information about the structure of gpA was obtained through the use of hybrid phages resulting from recombination between lambda and the related phage 21. lambda and 21 encode terminases that are analogous in structural organization and have ca. 60% sequence identity. In spite of these similarities, lambda and 21 terminases differ in specificity for DNA binding, subunit assembly, and prohead binding. A lambda-21 hybrid phage produces a terminase in which one of the subunits is chimeric and had recombinant specificities. In the work reported here; a new hybrid, lambda-21 hybrid 67, is characterized. lambda-21 hybrid 67 is the result of a crossover between lambda and 21 in the large subunit genes, such that the DNA from the left chromosome end is from 21, including cosB phi 21, the 1 gene, and the first 48 codons for the 2 gene. The rest of the hybrid 67 chromosome is lambda DNA, including 593 codons of the A gene. The chimeric gp2/A of hybrid 67 binds gp1 to form functional terminase.(ABSTRACT TRUNCATED AT 250 WORDS)

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Glutamate Dehydrogenase from Thermus thermophilus Is Activated by AMP and Leucine as a Complex with Catalytically Inactive Adenine Phosphoribosyltransferase Homolog

Journal of Bacteriology ◽

10.1128/jb.00710-18 ◽

2019 ◽

Vol 201 (14) ◽

Cited By ~ 1

Author(s):

Takeo Tomita ◽

Hajime Matsushita ◽

Ayako Yoshida ◽

Saori Kosono ◽

Minoru Yoshida ◽

...

Keyword(s):

Enzymatic Activity ◽

Glutamate Dehydrogenase ◽

Ternary Complex ◽

Thermus Thermophilus ◽

Thermophilic Bacterium ◽

Regulatory Subunit ◽

Binary Complex ◽

Bacterial Cells ◽

Adenine Phosphoribosyltransferase ◽

Content Type

ABSTRACT Glutamate dehydrogenase (GDH) from a thermophilic bacterium, Thermus thermophilus, is composed of two heterologous subunits, GdhA and GdhB. In the heterocomplex, GdhB acts as the catalytic subunit, whereas GdhA lacks enzymatic activity and acts as the regulatory subunit for activation by leucine. In the present study, we performed a pulldown assay using recombinant T. thermophilus, producing GdhA fused with a His tag at the N terminus, and found that TTC1249 (APRTh), which is annotated as adenine phosphoribosyltransferase but lacks the enzymatic activity, was copurified with GdhA. When GdhA, GdhB, and APRTh were coproduced in Escherichia coli cells, they were purified as a ternary complex. The ternary complex exhibited GDH activity that was activated by leucine, as observed for the GdhA-GdhB binary complex. Furthermore, AMP activated GDH activity of the ternary complex, whereas such activation was not observed for the GdhA-GdhB binary complex. This suggests that APRTh mediates the allosteric activation of GDH by AMP. The present study demonstrates the presence of complicated regulatory mechanisms of GDH mediated by multiple compounds to control the carbon-nitrogen balance in bacterial cells. IMPORTANCE GDH, which catalyzes the synthesis and degradation of glutamate using NAD(P)(H), is a widely distributed enzyme among all domains of life. Mammalian GDH is regulated allosterically by multiple metabolites, in which the antenna helix plays a key role to transmit the allosteric signals. In contrast, bacterial GDH was believed not to be regulated allosterically because it lacks the antenna helix. We previously reported that GDH from Thermus thermophilus (TtGDH), which is composed of two heterologous subunits, is activated by leucine. In the present study, we found that AMP activates TtGDH using a catalytically inactive APRTh as the sensory subunit. This suggests that T. thermophilus possesses a complicated regulatory mechanism of GDH to control carbon and nitrogen metabolism.

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Fluorescence Enhancement of Europium(III) Perchlorate by 2,2′-Dipyridyl on Bis(benzylsulfinyl)methane Complex

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.634-638.2462 ◽

2013 ◽

Vol 634-638 ◽

pp. 2462-2465

Author(s):

Wen Xian Li ◽

Bo Yang Ao ◽

Jing Zhang

Keyword(s):

Rare Earth ◽

Ternary Complex ◽

Fluorescence Enhancement ◽

Fluorescence Emission ◽

Europium Complex ◽

New Method ◽

Rare Earth Complexes ◽

Fluorescence Properties ◽

Binary Complex ◽

Fluorescence Lifetimes

A novel ligand with double sulfinyl groups, bis(benzylsulfinyl)methane L, was synthesized by a new method. Its novel ternary complex, has been synthesized [using L as the first ligand, and dipyridyl L' as the second ligand]. In order to study the effect of the second ligand on the fluorescence properties of rare-earth sulfoxide complex, a novel binary europium complex has been synthesized. Photoluminescent measurement showed that the first ligand L could efficiently transfer the energy to Eu (III) ions in the complex. Furthermore, the detailed luminescence analyses on the rare earth complexes indicated that the ternary Eu (III) complex manifested stronger fluorescence intensities, longer lifetimes, and higher fluorescence quantum efficiencies than the binary Eu (III) materials. The fluorescence emission intensities and fluorescence lifetimes of the ternary complex enhanced more obviously than the binary complex.

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Crystal structure of hexanoyl-CoA bound to β-ketoacyl reductase FabG4 of Mycobacterium tuberculosis

Biochemical Journal ◽

10.1042/bj20121107 ◽

2013 ◽

Vol 450 (1) ◽

pp. 127-139 ◽

Cited By ~ 27

Author(s):

Debajyoti Dutta ◽

Sudipta Bhattacharyya ◽

Amlan Roychowdhury ◽

Rupam Biswas ◽

Amit Kumar Das

Keyword(s):

Mycobacterium Tuberculosis ◽

Active Site ◽

Ternary Complex ◽

Catalytic Mechanism ◽

Acyl Carrier Protein ◽

Structural Data ◽

Acid Synthesis ◽

Binary Complex ◽

C Terminus ◽

Covalently Linked

FabGs, or β-oxoacyl reductases, are involved in fatty acid synthesis. The reaction entails NADPH/NADH-mediated conversion of β-oxoacyl-ACP (acyl-carrier protein) into β-hydroxyacyl-ACP. HMwFabGs (high-molecular-weight FabG) form a phylogenetically separate group of FabG enzymes. FabG4, an HMwFabG from Mycobacterium tuberculosis, contains two distinct domains, an N-terminal ‘flavodoxintype’ domain and a C-terminal oxoreductase domain. The catalytically active C-terminal domain utilizes NADH to reduce β-oxoacyl-CoA to β-hydroxyacyl-CoA. In the present study the crystal structures of the FabG4–NADH binary complex and the FabG4–NAD+–hexanoyl-CoA ternary complex have been determined to understand the substrate specificity and catalytic mechanism of FabG4. This is the first report to demonstrate how FabG4 interacts with its coenzyme NADH and hexanoyl-CoA that mimics an elongating fattyacyl chain covalently linked with CoA. Structural analysis shows that the binding of hexanoyl-CoA within the active site cavity of FabG significantly differs from that of the C16 fattyacyl substrate bound to mycobacterial FabI [InhA (enoyl-ACP reductase)]. The ternary complex reveals that both loop I and loop II interact with the phosphopantetheine moiety of CoA or ACP to align the covalently linked fattyacyl substrate near the active site. Structural data ACP inhibition studies indicate that FabG4 can accept both CoA- and ACP-based fattyacyl substrates. We have also shown that in the FabG4 dimer Arg146 and Arg445 of one monomer interact with the C-terminus of the second monomer to play pivotal role in substrate association and catalysis.

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Refined structure of the FKBP12–rapamycin–FRB ternary complex at 2.2 Å resolution

Acta Crystallographica Section D Biological Crystallography ◽

10.1107/s0907444998014747 ◽

1999 ◽

Vol 55 (4) ◽

pp. 736-744 ◽

Cited By ~ 78

Author(s):

Jun Liang ◽

Jungwon Choi ◽

Jon Clardy

Keyword(s):

Cell Cycle ◽

Biological Activity ◽

High Resolution ◽

Protein Interactions ◽

Ternary Complex ◽

Binary Complex ◽

Fk506 Binding Protein ◽

Cycle Arrest ◽

Protein Partners

The structure of the FKBP12–rapamycin–FRB ternary complex has now been refined at 2.2 Å resolution. The cell-cycle arrest agent rapamycin binds FK506-binding protein (FKBP12) and the FKBP12–rapamycin binding (FRB) domain of FKBP12–rapamycin associated protein (FRAP) simultaneously, and the inhibition of FRAP is responsible for rapamycin's biological activity. The conformation of rapamycin in the ternary complex is very similar to that observed in the FKBP12–rapamycin binary complex, with an r.m.s. difference of only 0.30 Å. However, a slight (9°) rotation repositions the FRB-binding face of rapamycin in the ternary complex. There are extensive rapamycin–protein interactions and relatively few interactions between the two protein partners FKBP12 and FRB, these interactions mainly involving residues in the 40s and 80s loops of FKBP12 and α1 and α4 of FRB. The high-resolution refinement has revealed the crucial role of several buried waters in the formation of the ternary complex.

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The binding of α-ketoglutarate in a binary complex and in a ternary complex with NADP+ by l-glutamate dehydrogenase

Biochimica et Biophysica Acta (BBA) - Enzymology ◽

10.1016/0005-2744(72)90104-0 ◽

1972 ◽

Vol 289 (1) ◽

pp. 28-36 ◽

Cited By ~ 30

Author(s):

D.G. Cross ◽

L.L. McGregor ◽

H.F. Fisher

Keyword(s):

Glutamate Dehydrogenase ◽

Ternary Complex ◽

Binary Complex

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FLUORESCENCE ENHANCEMENT OF Al3+– SODIUM MORIN–5–SULFONATE COMPLEX BY IMIDAZOLIUM IONIC LIQUID AND ITS APPLICATION IN DETERMINATION OF Al3+ IONS IN AN AQUEOUS SOLUTION

Jurnal Teknologi ◽

10.11113/jt.v81.12891 ◽

2019 ◽

Vol 81 (2) ◽

Cited By ~ 1

Author(s):

Syaza Atikah Nizar ◽

Nurul Syamimi Abdul Satar ◽

Shaik Azri Shaik Amar ◽

Fatin Hazirah Abdullah ◽

Faizatul Shimal Mehamod ◽

...

Keyword(s):

Aqueous Solution ◽

Ionic Liquid ◽

Acetic Acid ◽

Ternary Complex ◽

Fluorescence Enhancement ◽

Calibration Graph ◽

Fluorescence Signal ◽

Binary Complex ◽

Imidazolium Ionic Liquid

This study describes the preparation of sodium morin–5–sulfonate (NaMSA) as a new reagent for the determination of aluminium(III) (Al3+) ions based on the formation of a ternary complex. The complex consists of Al3+, NaMSA, and 1–Butyl–3–methylimidazolium hexafluorophosphate (BMIM–PF6). It was found that this method was sensitive compared to the binary complex of Al3+ and NaMSA. The ternary complex was excited at 420 nm, and the fluorescence signal was measured at 518 nm. Maximum fluorescence signal produced at pH 5.0 (acetic acid–acetate buffer), with 0.02% v/v BMIM–PF6 and 1.0 × 10-4 molL-1 of NaMSA. The calibration graph in linear up to 10 mgL-1 with the calculated detection limit of 0.017 mgL-1. Effect of foreign ions towards the ternary complex was also studied. Finally, the method was applied in the determination of Al3+ ions in water samples, and satisfactory results were obtained.

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Synthesis, Characterization and Luminescence of Europium, Terbium Complexes of 1,1'-(Pyridin-2,6-Diyl) Bis-3-P-Tolylpropane-1,3-Dione

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.306-307.228 ◽

2011 ◽

Vol 306-307 ◽

pp. 228-233

Author(s):

Jin Liang Zhu ◽

Sheng Li Liu ◽

Liang Cai Yu ◽

Tao Ouyang

Keyword(s):

Crystal Structure ◽

Nmr Spectroscopy ◽

Fluorescence Spectrum ◽

Ternary Complex ◽

Luminescent Properties ◽

Lanthanide Ions ◽

Binary Complex ◽

H Nmr ◽

Terbium Complexes ◽

Luminescent Compound

A new ligand, 1,1'-(pyridin-2,6-diyl)bis-3-p-tolylpropane-1,3-dione(H2L), was prepared and its crystal structure displays its enol isomer. The binary complex, Ln2L3·4H2O [Ln = Eu(III) and Tb(III)], were synthesized by the reaction of synthesized ligand (H2L) and Ln(III). With 1,10-phenanthroline (Phen) as the synergistic ligand, the ternary complex, Ln2L3(Phen)2 [Ln = Eu(III) and Tb(III)], were synthesized. The compounds were characterized by 1H-NMR spectroscopy, MS, IR, UV, DTA-TG and fluorescence spectrum. The luminescent properties of the ligand and its complexes were investigated and the results showed that the prepared ligand was a good luminescent compound, all complexes emitted characteristic luminescence of corresponding lanthanide ions and 1,10-phenanthroline as the synergistic ligand can obviously enhance the luminescence of Eu(III) in the ternary complex.

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The Endothelial Protein C Receptor Supports TF-VIIa-Xa Ternary Complex Signaling through Protease-Activated Receptors.

Blood ◽

10.1182/blood.v116.21.1144.1144 ◽

2010 ◽

Vol 116 (21) ◽

pp. 1144-1144

Author(s):

Jennifer Disse ◽

Helle Heibroch Petersen ◽

Katrine S. Larsen ◽

Egon Persson ◽

Naomi Esmon ◽

...

Keyword(s):

Protein C ◽

Ternary Complex ◽

Coagulation Factor ◽

Factor Vii ◽

Binary Complex ◽

Endothelial Protein C Receptor ◽

Free System ◽

Isolated Lung ◽

Complex Signaling ◽

Human And Mouse

Abstract Abstract 1144 Protease-activated receptor (PAR) signaling is closely linked to the cellular activation of the pro- and anticoagulant pathways. In contrast to thrombin that directly binds to and activates PAR1, other coagulation factors are dependent on co-receptors for efficient PAR cleavage. The endothelial protein C receptor (EPCR) is crucial for protein C (PC) activation by thrombomodulin-bound thrombin and supports signaling of activated PC through PAR1. EPCR may play additional roles by interacting with procoagulant proteases. EPCR binds the Gla-domain of human coagulation factor VII and conflicting reports exist about the role of EPCR as a receptor for FX. We studied the interaction of murine soluble EPCR extracellular domain (sEPCR) under physiological concentrations of divalent cations Ca2+/Mg2+. BIAcore measurements showed that murine sEPCR bound both human and mouse complexes of soluble tissue factor (TF) with FVIIa, as well as human FX. In a lipid free system measuring only protein-protein interactions, amidolytic activity of soluble TF-FVIIa was not changed by sEPCR. However, sEPCR dose dependently inhibited FX activation by both human and mouse soluble TF-FVIIa, indicating that sEPCR interacted with the TF-FVIIa-FX extrinsic activation complex. On human cells, TF forms two signaling complexes, the non-coagulant TF-FVIIa binary complex that activates PAR2 and the ternary TF-FVIIa-FXa complex that signals through PAR1 or PAR2. Overexpression of PAR2 in HUVECs resulted in PAR2 activation by activated PC, and in these transduced cells TF-FVIIa-FXa ternary complex signaling was inhibited by antibody blockade of EPCR. Human HaCaT keratinocytes constitutively synthesize TF and represent a well characterized model of TF binary and ternary complex signaling through PAR2. FACS analysis showed that EPCR was expressed on the cell surface and antibody blockade of EPCR prevented TF-FVIIa-Xa ternary, but not TF-FVIIa binary complex signaling. Mutation of FVIIa Gla residues had no effect on ternary complex signaling, indicating a primary interaction of EPCR with FX/FXa. To expand these studies to murine cells that constitutively express EPCR and TF, we isolated lung smooth muscle cells (SMC) from wild-type, PAR1-/-, PAR2-/-, and EPCRlow mice. Stimulation of SMC with thrombin, high concentrations of FXa, or FVIIa/FX, but not FVIIa alone induced PAR1-dependent signaling. While thrombin signaling was unaltered, EPCRlow SMC showed no response to the ternary complex measured by ERK1/2 phosphorylation. In order to exclude indirect effects on SMC phenotypes due to prolonged EPCR-deficiency in vivo, we further deleted EPCR in vitro by adenoviral transduction with cre recombinase from EPCRfloxflox SMC or blocked EPCR function with antibodies to murine EPCR. Both approaches inhibited TF ternary complex, but not thrombin signaling. These results show that EPCR interacts with the TF coagulation initiation complex to enable specifically ternary complex signaling and suggest that EPCR may play a role in regulating the biology of TF expressing extravascular and vessel wall cells that are exposed to limited concentrations of FVIIa and FX provided by ectopic synthesis or vascular leakage. Disclosures: Heibroch Petersen: Novo Nordisk: Employment. Persson:Novo Nordisk: Employment. Petersen:Novo Nordisk: Employment. Ruf:Novo Nordisk: Research Funding.

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NADP-Dependent Aldehyde Dehydrogenase from Archaeon Pyrobaculum sp.1860: Structural and Functional Features

Archaea ◽

10.1155/2016/9127857 ◽

2016 ◽

Vol 2016 ◽

pp. 1-14

Author(s):

Ekaterina Yu. Bezsudnova ◽

Tatiana E. Petrova ◽

Natalia V. Artemova ◽

Konstantin M. Boyko ◽

Ivan G. Shabalin ◽

...

Keyword(s):

Aldehyde Dehydrogenase ◽

Active Sites ◽

Ternary Complex ◽

Binary Complex ◽

Relay Systems ◽

Binary Complexes ◽

Proton Relay ◽

Functional Features

We present the functional and structural characterization of the first archaeal thermostable NADP-dependent aldehyde dehydrogenase AlDHPyr1147. In vitro, AlDHPyr1147 catalyzes the irreversible oxidation of short aliphatic aldehydes at 60–85°С, and the affinity of AlDHPyr1147 to the NADP+ at 60°С is comparable to that for mesophilic analogues at 25°С. We determined the structures of the apo form of AlDHPyr1147 (3.04 Å resolution), three binary complexes with the coenzyme (1.90, 2.06, and 2.19 Å), and the ternary complex with the coenzyme and isobutyraldehyde as a substrate (2.66 Å). The nicotinamide moiety of the coenzyme is disordered in two binary complexes, while it is ordered in the ternary complex, as well as in the binary complex obtained after additional soaking with the substrate. AlDHPyr1147 structures demonstrate the strengthening of the dimeric contact (as compared with the analogues) and the concerted conformational flexibility of catalytic Cys287 and Glu253, as well as Leu254 and the nicotinamide moiety of the coenzyme. A comparison of the active sites of AlDHPyr1147 and dehydrogenases characterized earlier suggests that proton relay systems, which were previously proposed for dehydrogenases of this family, are blocked in AlDHPyr1147, and the proton release in the latter can occur through the substrate channel.

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