Crystallization of protein–protein complexes

Crystallizing protein–protein complexes remains a rate-limiting step in their structure characterization. Crystallization conditions for the known protein–protein complexes have been surveyed in both the Protein Data Bank and the BMCD database. Compared with non-complexed proteins, crystallization conditions for protein–protein complexes are less diverse and heavily favor (71%versus27%) polyethylene glycols (PEG) rather than ammonium sulfate or other high-salt crystallization conditions. The results suggest that the stability of protein complexes limits their available crystallization configuration space. Based on the survey, a set of sparse-matrix screen conditions was designed.

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Tertiary amine-catalyzed generation of chlorine dioxide from hypochlorous acid and chlorite ions

Wood Science and Technology ◽

10.1007/s00226-020-01247-5 ◽

2020 ◽

Author(s):

Estefania Isaza Ferro ◽

Jordan Perrin ◽

Owain George John Dawson ◽

Tapani Vuorinen

Keyword(s):

Chlorine Dioxide ◽

Tertiary Amine ◽

Hypochlorous Acid ◽

Suitable Candidate ◽

Rate Limiting Step ◽

Limiting Step ◽

Chlorite Ions ◽

The Stability ◽

Rate Limiting

AbstractThe reaction between hypochlorous acid and chlorite ions is the rate limiting step for in situ chlorine dioxide regeneration. The possibility of increasing the speed of this reaction was analyzed by the addition of tertiary amine catalysts in the system at pH 5. Two amines were tested, DABCO (1,4-diazabicyclo[2.2.2]octane) and its derivative CEM-DABCO (1-carboethoxymethyl-1-azonia-4-aza-bicyclo[2.2.2]octane chloride). The stability of the catalysts in the presence of both reagents and chlorine dioxide was measured, with CEM-DABCO showing to be highly stable with the mentioned chlorine species, whereas DABCO was rapidly degraded by chlorine dioxide. Hence, CEM-DABCO was chosen as a suitable candidate to catalyze the reaction of hypochlorous acid with chlorite ions and it significantly increased the speed of this reaction even at low catalyst dosages. This research opens the door to a faster regeneration of chlorine dioxide and an improved efficiency in chlorine dioxide treatments.

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Charge screening by cations affects the conformation of the mitochondrial inner membrane. A study of exogenous NAD(P)H oxidation in plant mitochondria

Biochemical Journal ◽

10.1042/bj1950583 ◽

1981 ◽

Vol 195 (3) ◽

pp. 583-588 ◽

Cited By ~ 26

Author(s):

I M Møller ◽

J M Palmer

Keyword(s):

Electron Transport ◽

Protein Complexes ◽

Plant Mitochondria ◽

Lateral Diffusion ◽

Jerusalem Artichoke ◽

Inner Membrane ◽

Mitochondrial Inner Membrane ◽

Rate Limiting Step ◽

Limiting Step ◽

Rate Limiting

Cations caused a decrease in the apparent Km and an increase in the Vmax. for the oxidation of exogenous NADH by both Jerusalem-artichoke (Helianthus tuberosus) and Arum maculatum (cuckoo-pint) mitochondria prepared and suspended in a low-cation medium (approximately or equal to 1 mM-K+). In Arum mitochondria the addition of cations caused a much greater stimulation of the oxidation of NAD(P)H via the cytochrome oxidase pathway than via the alternative, antimycin-insensitive, pathway. This shows that cations affected a rate-limiting step in the electron-transport chain at or beyond ubiquinone, the branch-point of electron transport in plant mitochondria. The effects were only dependent on the valency of the cation (efficiency C3+ greater than C2+ greater than C+) and not on its chemical nature, which is consistent with the theory of the diffuse layer. The results are interpreted to show that the screening of fixed negative membrane changes on lipids and protein complexes causes a conformational change in the mitochondrial inner membrane, leading to a change in a rate-limiting step of NAD(P)H oxidation. More specifically, it is proposed that screening removes electrostatic restrictions on lateral diffusion and thus accelerates diffusion-limited steps in electron transport.

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Influence of Cs Loading on Pt/m-ZrO2 Water–Gas Shift Catalysts

Catalysts ◽

10.3390/catal11050570 ◽

2021 ◽

Vol 11 (5) ◽

pp. 570

Author(s):

Zahra Rajabi ◽

Michela Martinelli ◽

Caleb D. Watson ◽

Donald C. Cronauer ◽

A. Jeremy Kropf ◽

...

Keyword(s):

Alkali Metals ◽

Fixed Bed ◽

Fixed Bed Reactor ◽

Co2 Removal ◽

Electronic Density ◽

Rate Limiting Step ◽

Limiting Step ◽

The Stability ◽

Rate Limiting ◽

Co2 Decomposition

Certain alkali metals (Na, K) at targeted loadings have been shown in recent decades to significantly promote the LT-WGS reaction. This occurs at alkali doping levels where a redshift in the C-H band of formate occurs, indicating electronic weakening of the bond. The C-H bond breaking of formate is the proposed rate-limiting step of the formate associative mechanism, lending support to the occurrence of this mechanism in H2-rich environments of the LT-WGS stage of fuel processors. Continuing in this vein of research, 2%Pt/m-ZrO2 was promoted with various levels of Cs in order to explore its influence on the rate of formate intermediate decomposition, as well as that of LT-WGS in a fixed bed reactor. In situ DRIFTS experiments revealed that Cs promoter loadings of 3.87% to 7.22% resulted in significant acceleration of the forward formate decomposition in steam at 130 °C. Of all of the alkali metals tested to date, the redshift in the formate ν(CH) band with the incorporation of Cs was the greatest. XANES difference experiments at the Pt L2 and L3 edges indicated that the electronic effect was not likely due to an enrichment of electronic density on Pt. CO2 TPD experiments revealed that, unlike Na and K promoters, Cs behaves more like Rb in that the decomposition of the second intermediate in LT-WGS, carbonate species, is hindered due to (1) increased basicity of Cs, (2) the tendency of Cs to cover Pt sites that facilitate CO2 decomposition, and (3) the tendency of Cs to increase Pt particle size as shown by EXAFS results, resulting in fewer Pt sites that facilitate CO2 decomposition. As such, the LT-WGS rate was hindered overall and the rate-limiting step shifted to carbonate decomposition (CO2 removal). Like its Rb counterpart, low levels of added Cs (e.g., 0.72%Cs) were found to improve the stability of the catalyst relative to the unpromoted catalyst; the stability comparison was made at similar CO conversion level as well as similar space velocity.

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A beginner’s guide to macromolecular crystallization

The Biochemist ◽

10.1042/bio_2020_108 ◽

2021 ◽

Vol 43 (1) ◽

pp. 36-43

Author(s):

Fabrice Gorrec

Keyword(s):

Crystal Nucleation ◽

Trial And Error ◽

Bioinformatics Analyses ◽

X Ray ◽

X Ray Crystallography ◽

Rate Limiting Step ◽

Limiting Step ◽

Crystallization Conditions ◽

Macromolecular Crystallization ◽

Rate Limiting

Obtaining diffraction-quality crystals is currently the rate-limiting step in macromolecular X-ray crystallography of proteins, DNA, RNA or their complexes, in the vast majority of cases. Since each sample has different and specific characteristics – which is the reason for wanting to study every single one of them in the first place – crystallization conditions cannot be predicted. Hence, researchers must enable crystal nucleation and growth through experimentation and screening. The size, shape and surface of the sample or complexes of interest are often altered through genetic and biochemical manipulation to facilitate crystallization, based on bioinformatics analyses and trial and error. Pure samples are trialled against a very broad range of crystallization conditions. The currently predominant method to achieve crystallization is sitting drop vapour diffusion with nanolitre-class robotic liquid handlers. Once initial screening yields crystals, further optimization experiments are usually required to obtain larger and diffraction-quality crystals.

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Solid products and rate-limiting step in the thermal half decomposition of natural dolomite in a CO2(g) atmosphere

Thermochimica Acta ◽

10.1016/s0040-6031(02)00603-2 ◽

2003 ◽

Cited By ~ 1

Author(s):

D Beruto

Keyword(s):

Rate Limiting Step ◽

Limiting Step ◽

Rate Limiting

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Ornithine Decarboxylase Activity in Cultured Endothelial Cells Stimulated by Serum, Thrombin and Serotonin

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646709 ◽

1978 ◽

Vol 39 (02) ◽

pp. 496-503 ◽

Cited By ~ 8

Author(s):

P A D’Amore ◽

H B Hechtman ◽

D Shepro

Keyword(s):

Endothelial Cells ◽

Ornithine Decarboxylase ◽

Decarboxylase Activity ◽

Aortic Endothelial Cells ◽

Ornithine Decarboxylase Activity ◽

Rate Limiting Step ◽

Bovine Aortic Endothelial Cells ◽

Limiting Step ◽

Rate Limiting ◽

Serum Serotonin

SummaryOrnithine decarboxylase (ODC) activity, the rate-limiting step in the synthesis of polyamines, can be demonstrated in cultured, bovine, aortic endothelial cells (EC). Serum, serotonin and thrombin produce a rise in ODC activity. The serotonin-induced ODC activity is significantly blocked by imipramine (10-5 M) or Lilly 11 0140 (10-6M). Preincubation of EC with these blockers together almost completely depresses the 5-HT-stimulated ODC activity. These observations suggest a manner by which platelets may maintain EC structural and metabolic soundness.

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