Pyrococcus furiosus ferredoxin is a functional dimer.

M N Hasan; P L Hagedoorn; W R Hagen

doi:10.1016/s0014-5793(02)03554-8

Home/Archive/PubMed

PubMed · 12417337

Pyrococcus furiosus ferredoxin is a functional dimer.

M N Hasan·P L Hagedoorn·W R Hagen

View original source ↗Export metadata ↓

Abstract

Pyrococcus furiosus ferredoxin is subject to a monomer/dimer equilibrium as a function of ionic strength. At physiological ionic strength, approximately 0.35 M NaCl, the protein is very predominantly homodimer. The monomeric form exhibits impaired electron transfer on glassy carbon; it also has a decreased S=3/2 over S=1/2 ratio as shown by electron paramagnetic resonance spectroscopy. Even following sterilization at 121 degrees C the dimer is stable in denaturing gel electrophoresis.

Explore related subjects

Chromatography, Gel Dimerization Electron Spin Resonance Spectroscopy Electron Transport Electrophoresis, Polyacrylamide Gel Ferredoxins Osmolar Concentration Pyrococcus furiosus Journal Article

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

M N Hasan, P L Hagedoorn, W R Hagen. 2002-11-06. Pyrococcus furiosus ferredoxin is a functional dimer.. https://doi.org/10.1016/s0014-5793(02)03554-8

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2007

The influence of grafted polymer architecture and fluid hydrodynamics on protein separation by entropic interaction chromatography.

Entropic interaction chromatography (EIC) provides efficient size-based separation of protein mixtures through the entropy change associated with solute partitioning into a layer of hydrophilic homopolymer that has been end-grafted within the pores of a macroporous chromatography support. In this work, surface-initiated atom-transfer radical polymerization (ATRP) is used to prepare a library of EIC stationary phases covering a wide range of grafted-chain densities and molecular weights. Exhaustive chain cleavage and analysis by saponification and GPC-MALLS, respectively, show that the new ATRP synthesis procedure allows for excellent control over graft molecular weight and polydispersity. The method is used to prepare high-density grafts (up to 0.164 +/- 0.005 chains/nm(2)) that extend the range of EIC applications to include efficient buffer-exchange and desalting of protein preparations. Reducing the graft density allows for greater partitioning of high molecular weight solutes, extending the linear range of the selectivity curve. Increasing graft molecular weight also alters selectivity, but more directly affects column capacity by increasing the volume of the grafted layer. Protein partitioning in high-density EIC columns is found to decrease with mobile-phase velocity (u). Although solute mass transfer resistances leading to an increase in plate height can explain this effect, pressure drop data across the column are indicative of weak convective flow through at least a fraction of the grafted architecture. Modeling of the grafted brush properties in the presence of solvent flow by subjecting a self-consistent-field theory representation of the brush to a viscous shear force predicts that the grafted chains will tilt and elongate in the direction of flow. The shear force may therefore act to reduce the number of conformations available to chains, increasing their rigidity without significantly altering the thickness of the grafted layer. A reduction in protein partitioning is then predicted when the dependence on u of the solute entropy loss is stronger than that of the grafted polymer, a condition met at high graft densities.

Chromatography, Gel↗

PubMed2007

Effects of gamma radiation on beta-lactoglobulin: oligomerization and aggregation.

The conformational changes and aggregation process of beta-lactoglobulin (beta-LG) subjected to gamma irradiation are presented. Beta-LG in solutions of different protein concentrations (3 and 10 mg/ml) and in solid state with different water activities (a(w)) (0.22; 0.53; 0.74) was irradiated using a Cobalt-60 radiation source at dose level of 1-50 kGy. Small-angle X-ray scattering (SAXS) was used to study the conformational changes of beta-LG due to the irradiation treatment. The irradiated protein was also examined by high performance size exclusion chromatography (HPSEC) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing and reducing conditions and fluorescence. SAXS analysis showed that the structural conformation of irradiated beta-LG in solid state at different a(w) and dose level was essentially the same as the nonirradiated beta-LG. The scattering data also showed that the irradiation of beta-LG in solution promoted the formation of oligomers. Interestingly, from the data analysis and model building, it could be shown that the formed oligomers are linear molecules, built by linear combinations of beta-LG dimers (tetramers, hexamers, etc). The formation of oligomers was also evidenced by SDS-PAGE analysis and HPSEC chromatograms, in which products with higher molecular mass than that of the dimeric beta-LG were detected. Formation of intermolecular cross-linking between tyrosyl radicals are proposed to be at least partially responsible for this occurrence. From the results it could be shown that the samples irradiated in solution presented some conformational changes under gamma irradiation, resulting in well ordered oligomers and aggregates formed by cross-linking of beta-LG dimers subunits, while the samples irradiated in the solid state were not modified.

Chromatography, Gel↗

PubMed2007

Comparison of two gel filtration chromatographic methods for the purification of Lily symptomless virus.

Lily symptomless virus (LSV) occurs frequently in many Lilium species worldwide and often causes developmental abnormalities such as a smaller flower and lower bulb yield. In this study, two moderate and efficient gel filtration chromatography (GFC) methods was compared, these two techniques were, respectively, based on Superdex-200 HR and Sephacryl S-1000 SF. The products purified by the two methods were then characterized by measurements with UV-spectrophotometer, reverse transcriptase (RT)-PCR, transmission electron microscope (TEM), polyacrylamide gel electrophoresis (PAGE), Western blotting and matrix assisted laser desorption-ionisation/time-of-flight mass spectrometry (MALDI-TOF-MS). The final yield of purified LSV by the Superdex-200 HR GFC was 9.4 mg from 50 g of fresh infected tissues of Lanzhou lily. However, from the same amount tissues, only 5.6 mg of LSV were obtained by using Sephacryl S-1000 SF GFC. The Superdex-200 HR method was thus shown to be more suitable for the purification of LSV than the Sephacryl S-1000 SF GFC. The Superdex-200 HR method does not require costly equipment for density centrifugation and ultracentrifugation. Furthermore, it can provide an economical and efficient way to obtain purified products for the preparation of antibodies for serological diagnosis or LSV infection and related investigations.

Chromatography, Gel↗