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Dynamic Light Scattering Used for Allosteric Transition Characterization

Malvern Panalytical
23 Jul 2012
Tesni Perry
Administrator / Office Personnel

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Using the Zetasizer µV dynamic light scattering (DLS) system from Malvern Instruments, Professor Mario L. Calcagno and his team at the Biochemistry Department of the Faculty of Medicine, Universidad Nacional Autónoma de México (UNAM, the National Autonomous University of Mexico) have been able to distinguish allosteric transition [structural rearrangements] in a single E. coli protein.

Characterizing protein allostery is challenging because of its sensitivity to experimental conditions, however, a quantitative description of allosteric transition is important in understanding and controlling metabolic and other biochemical processes.

“We use the Zetasizer µV to characterize the size of proteins and how they interact to produce multimeric forms or even supramolecular arrangements of the protein such as viral capsids [shells],” said Dr Ismael Bustos-Jaimes, from Laboratory of Physical Chemistry and Protein Engineering at UNAM commented “The Zetasizer µV allows us to work with sizes in the range of 1 nm to 500 nm and follow each assembly and stability parameter, such as pH, temperature and ionic strength, guiding us to the optimal production conditions for these virus-like particles.”

“The sensitivity of the Zetasizer µV has additionally allowed my colleague, Prof Calcagno, to analyze allosteric transitions” explained Dr Bustos-Jaimes. “The size of the hexameric glucosamine-6-phosphate deaminase protein from E. coli changes its shape to a more compact form upon allosteric-activator binding, and this change can be measured.”

“The Zetasizer µV software is user-friendly and in addition to measuring particle size it delivers information about the quality of sample preparation. This is very important when you work with molecules which are prone to uncontrolled aggregation,” said Dr Bustos-Jaimes.

The UNAM team studies allosteric transitions and the assembly of virus-like particles (VLPs) for use in diagnostics and disease control. VLPs are biological nanoparticles that resemble natural viruses but contain no genetic material. As non-infective agents, they are suitable for use in the analysis of viral infection mechanisms, vaccine production, tissue-specific drug delivery and as biological nanomaterials.

Malvern, Malvern Instruments and Zetasizer are registered trademarks of Malvern Instruments Ltd

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Size Exclusion ChromatographySize exclusion chromatography (SEC) is a liquid chromatographic technique used to separate and identify molecules based on their size, including polymers, proteins, nanoparticles & nucleic acid. SEC is further subdivided into gel filtration chromatography (GFC) and gel permeation chromatography (GPC) when an aqueous or organic mobile phase is used respectively. HPLC-based SEC systems and columns are available for analytical separation alongside SEC spin columns or filters to prep samples for many protocols. Find the best size exclusion chromatography equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.MicrobiologyMicrobiology is the study of microorganisms including protists, prokaryotes, fungi, and, often, viruses. Microorganisms are a useful research tool as genetic vectors and, in immunology, for antibiotic susceptibility testing, cellular biology and genetics. Microorganisms commonly grow readily in incubators with microbial culture media; this can contain chromogenic supplements to differentiate between cell lines. Estimate your culture’s density of microorganisms with colony counters, or screen and select colonies for desirable clones with automated colony pickers. Additionally, equipment is available to monitor environments for the presence of microbes and identify with microbial identification instruments. Find the best microbiology products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Particle CharacterizationParticle characterization instruments are used to determine particle size distribution, shape, surface area, zeta potential, density and porosity of particles and materials. Multiple tecchniques are available for determining particle size, shape and count including dynamic light scattering (DLS), laser diffraction, electrozone (Coulter technique), imaging particle analysis and single particle optical sensing. Determine the density of your material with a gas pycnometer or examine its surface area and porosity with gas adsorption analyzers and mercury porosimeters. Find the best particle characterization instruments in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Viral Infection MechanicsVLPDynamic Light ScatteringDynamic light scattering is used to measure the size distribution of particles in suspension by analyzing the way light scatters off them. It is commonly used in materials science, nanotechnology, and protein characterization. Browse our peer-reviewed product directory to find the best dynamic light scattering systems, compare products, check reviews, and get pricing directly from manufacturers.

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