Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

Neutrons observe vitamin B6-dependent enzyme activity useful for drug development

17.10.2017

Scientists at the Department of Energy's Oak Ridge National Laboratory have performed neutron structural analysis of a vitamin B6-dependent protein, potentially opening avenues for new antibiotics and drugs to battle diseases such as drug-resistant tuberculosis, malaria and diabetes.

Specifically, the team used neutron crystallography to study the location of hydrogen atoms in aspartate aminotransferase, or AAT, an enzyme vital to the metabolism of certain amino acids.


An ORNL-led team used neutrons to observe the AAT enzyme, a vitamin B6-dependent protein, and found that the chemical reaction occurred only in one active site. Nuclear scattering length density maps (colored mesh) highlight the positions of critical hydrogen atoms, including a low-barrier hydrogen bond (magenta mesh) not thought to exist in AAT, which may be crucial for catalysis.

Credit: Jill Hemman and Andrey Kovalevsky /Oak Ridge National Laboratory, US Dept. of Energy

"We visualized the first neutron structure of a vitamin B6 enzyme that belongs to a large protein family with hundreds of members that exist in nature," said ORNL's Andrey Kovalevsky, a senior co-author of the study, which was published in Nature Communications.

Vitamin B6-dependent proteins are part of a diverse group of enzymes that conduct over a hundred different chemical reactions in cells. The enzymes are of interest to biomedical, as well as bioenergy, researchers because of their role in metabolizing amino acids and other cell nutrients.

"These enzymes are unique in that each one performs a specific chemical reaction with exquisite accuracy, while suppressing other viable chemical transformations," Kovalevsky said. "How they accomplish this is not well understood, but it is of great significance for drug design."

The team's previous research predicted that hydrogen atoms move in and around the enzyme's active site, where the chemical reaction takes place, indicating that the hydrogen atoms' positioning controls the reaction type. Knowing the precise location of hydrogen atoms can explain why the behavior of these enzymes is so specific, but hydrogen is hard to detect with standard methods such as X-ray crystallography.

To directly determine the positions of hydrogen atoms within AAT, the ORNL-led team turned to neutron diffraction techniques. The researchers exposed delicate protein crystals to neutrons using the IMAGINE beamline at ORNL's High Flux Isotope Reactor and the LADI-III beamline at the Institut Laue-Langevin in Grenoble, France.

Surprisingly, the team observed a reaction within one AAT protein biomolecule while another AAT biomolecule was unchanged, providing a before-and-after perspective of the enzyme-catalyzed chemical reaction.

"The data revealed that in one of the enzyme's biomolecular structures the covalent bonds reorganized after a chemical reaction occurred in the active site and, in another, the reaction had not taken place," Kovalevsky said. "Essentially, we were able to obtain two structures in one crystal, which has never been done before for any protein using neutrons."

With this knowledge, the team will run molecular simulations to determine the hydrogen atoms' specific behavior when interacting with the enzyme. The results could be useful in guiding the future design of novel medicines against multidrug-resistant tuberculosis, malaria, diabetes and antibiotic-resistant bacteria.

"This study highlights how neutrons are an unrivaled probe for identifying the location of hydrogen atoms in biological systems, providing us with an unprecedented level of structural detail for this important enzyme," LADI-III beamline scientist Matthew Blakeley said.

###

The neutron crystallography study, titled "Direct visualization of critical hydrogen atoms in a pyridoxal 5'- phosphate enzyme," included co-authors Steven Dajnowicz of ORNL and the University of Toledo; ORNL's Andrey Kovalevsky, Jerry Parks, Ryne C. Johnston and Kevin L. Weiss; Matthew P. Blakeley of the Institut Laue Langevin; David A. Keen of Rutherford Appleton Laboratory; Oksana Gerlits of the University of Tennessee, and Timothy C. Mueser of the University of Toledo.

Steven Dajnowicz, the paper's lead co-author and a doctoral student at the University of Toledo, works in ORNL's Biology and Soft Matter Division as part of ORNL's Graduate Opportunities (GO!) student program. Mueser is Dajnowicz' advisor and a senior co-author of the paper.

This project was funded by DOE's Office of Science and used resources at ORNL's Center for Structural Molecular Biology. Neutron crystallography work was performed at ORNL's HFIR, a DOE Office of Science User Facility, and at the Institut Laue-Langevin. This work also used ORNL's Compute and Data Environment for Science for quantum mechanical calculations. This project was also supported in part by the Center for the Advancement of Science in Space and the National Aeronautics and Space Administration.

ORNL is managed by UT-Battelle for the Department of Energy's Office of Science, the single largest supporter of basic research in the physical sciences in the United States. DOE's Office of Science is working to address some of the most pressing challenges of our time. For more information, please visit http://science.energy.gov.

Media Contact

Sara Shoemaker
shoemakerms@ornl.gov
865-576-9219

 @ORNL

http://www.ornl.gov 

Sara Shoemaker | EurekAlert!

More articles from Life Sciences:

nachricht Solving the efficiency of Gram-negative bacteria
22.03.2019 | Harvard University

nachricht Bacteria bide their time when antibiotics attack
22.03.2019 | Rice University

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: The taming of the light screw

DESY and MPSD scientists create high-order harmonics from solids with controlled polarization states, taking advantage of both crystal symmetry and attosecond electronic dynamics. The newly demonstrated technique might find intriguing applications in petahertz electronics and for spectroscopic studies of novel quantum materials.

The nonlinear process of high-order harmonic generation (HHG) in gases is one of the cornerstones of attosecond science (an attosecond is a billionth of a...

Im Focus: Magnetic micro-boats

Nano- and microtechnology are promising candidates not only for medical applications such as drug delivery but also for the creation of little robots or flexible integrated sensors. Scientists from the Max Planck Institute for Polymer Research (MPI-P) have created magnetic microparticles, with a newly developed method, that could pave the way for building micro-motors or guiding drugs in the human body to a target, like a tumor. The preparation of such structures as well as their remote-control can be regulated using magnetic fields and therefore can find application in an array of domains.

The magnetic properties of a material control how this material responds to the presence of a magnetic field. Iron oxide is the main component of rust but also...

Im Focus: Self-healing coating made of corn starch makes small scratches disappear through heat

Due to the special arrangement of its molecules, a new coating made of corn starch is able to repair small scratches by itself through heat: The cross-linking via ring-shaped molecules makes the material mobile, so that it compensates for the scratches and these disappear again.

Superficial micro-scratches on the car body or on other high-gloss surfaces are harmless, but annoying. Especially in the luxury segment such surfaces are...

Im Focus: Stellar cartography

The Potsdam Echelle Polarimetric and Spectroscopic Instrument (PEPSI) at the Large Binocular Telescope (LBT) in Arizona released its first image of the surface magnetic field of another star. In a paper in the European journal Astronomy & Astrophysics, the PEPSI team presents a Zeeman- Doppler-Image of the surface of the magnetically active star II Pegasi.

A special technique allows astronomers to resolve the surfaces of faraway stars. Those are otherwise only seen as point sources, even in the largest telescopes...

Im Focus: Heading towards a tsunami of light

Researchers at Chalmers University of Technology and the University of Gothenburg, Sweden, have proposed a way to create a completely new source of radiation. Ultra-intense light pulses consist of the motion of a single wave and can be described as a tsunami of light. The strong wave can be used to study interactions between matter and light in a unique way. Their research is now published in the scientific journal Physical Review Letters.

"This source of radiation lets us look at reality through a new angle - it is like twisting a mirror and discovering something completely different," says...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

VideoLinks
Industry & Economy
Event News

International Modelica Conference with 330 visitors from 21 countries at OTH Regensburg

11.03.2019 | Event News

Selection Completed: 580 Young Scientists from 88 Countries at the Lindau Nobel Laureate Meeting

01.03.2019 | Event News

LightMAT 2019 – 3rd International Conference on Light Materials – Science and Technology

28.02.2019 | Event News

 
Latest News

Solving the efficiency of Gram-negative bacteria

22.03.2019 | Life Sciences

Bacteria bide their time when antibiotics attack

22.03.2019 | Life Sciences

Open source software helps researchers extract key insights from huge sensor datasets

22.03.2019 | Information Technology

VideoLinks
Science & Research
Overview of more VideoLinks >>>