LACTIC DEHYDROGENASE OF THE MAMMALIAN ERYTHROCYTE: III. THE FORMATION OF A COMPLEX OF THE ENZYME WITH DPN AND LACTATE

1958 ◽  
Vol 36 (1) ◽  
pp. 1093-1098 ◽  
Author(s):  
Paul Ottolenghi ◽  
Orville F. Denstedt
Keyword(s):  
Ternary Complex ◽  
Kinetic Studies ◽  
Lactic Dehydrogenase ◽  
Binary Complex ◽  
Hydrogen Ions ◽  
Mammalian Erythrocyte

In the transfer of hydrogen ions and electrons from lactate to DPN by lactic dehydrogenase the lactate is capable of forming a complex only with the binary complex of enzyme and coenzyme to form an active ternary complex. The results of kinetic studies indicate that the site for the binding of lactate is on the DPN molecule itself.

LACTIC DEHYDROGENASE OF THE MAMMALIAN ERYTHROCYTE: III. THE FORMATION OF A COMPLEX OF THE ENZYME WITH DPN AND LACTATE

1958 ◽  
Vol 36 (10) ◽  
pp. 1093-1098 ◽  
Author(s):  
Paul Ottolenghi ◽  
Orville F. Denstedt
Keyword(s):  
Ternary Complex ◽  
Kinetic Studies ◽  
Lactic Dehydrogenase ◽  
Binary Complex ◽  
Hydrogen Ions ◽  
Mammalian Erythrocyte

In the transfer of hydrogen ions and electrons from lactate to DPN by lactic dehydrogenase the lactate is capable of forming a complex only with the binary complex of enzyme and coenzyme to form an active ternary complex. The results of kinetic studies indicate that the site for the binding of lactate is on the DPN molecule itself.

scholarly journals A larger number of L chains (Tac) enhance the association rate of interleukin 2 to the high affinity site of the interleukin 2 receptor.

1988 ◽  
Vol 168 (5) ◽  
pp. 1563-1572 ◽  
Author(s):  
Y Saito ◽  
H Sabe ◽  
N Suzuki ◽  
S Kondo ◽  
T Ogura ◽  
...  
Keyword(s):  
Ternary Complex ◽  
Interleukin 2 ◽  
Kinetic Studies ◽  
Complex Model ◽  
Fixed Number ◽  
Binary Complex ◽  
Conversion Model ◽  
Reaction Rate Constants ◽  
High Affinity ◽  
High Affinity Receptor

The IL-2-R is composed of at least two proteins, that is, a 55-kD protein (p55, the L chain, or Tac) and a 75-kD protein (p75, the H chain, or converter). The high affinity binding of IL-2 results in the formation of the ternary complex consisting of IL-2, and the L and H chains. To distinguish the affinity conversion model and the binary complex model we have carried out kinetic studies on the IL-2 binding to the high affinity IL-2-R on T lymphocytes expressing various numbers of L chains and a relatively constant number of H chains. We found that expression of a larger number of L chains accelerated the association of IL-2 to the high affinity receptor. The results are not compatible with the binary complex model that assumes a fixed number of high affinity sites determined by the numbers of a limiting chain. Instead, the results are consistent with the prediction of the affinity conversion model that assumes association of IL-2 to the L chain as the first step of the ternary complex formation and they indicate that the possible role of excess L chains is to accelerate the formation of the ternary complex. The reaction rate constants calculated from the affinity conversion model were reasonably constant.

scholarly journals Domains for protein-protein interactions at the N and C termini of the large subunit of bacteriophage lambda terminase.

Genetics ◽  
1988 ◽  
Vol 119 (3) ◽  
pp. 477-484
Author(s):  
W F Wu ◽  
S Christiansen ◽  
M Feiss
Keyword(s):  
Protein Interactions ◽  
Ternary Complex ◽  
Large Subunit ◽  
Small Subunit ◽  
Functional Domain ◽  
Binary Complex ◽  
Protein Protein Interactions ◽  
Related Phage ◽  
Carboxy Terminal ◽  
Lambda Dna

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)

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

2019 ◽  
Vol 201 (14) ◽  
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.

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

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.

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

2013 ◽  
Vol 450 (1) ◽  
pp. 127-139 ◽  
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.

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

1999 ◽  
Vol 55 (4) ◽  
pp. 736-744 ◽  
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.

scholarly journals 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

2019 ◽  
Vol 81 (2) ◽  
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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