Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

Scientists crack code of critical bacterial defense mechanism

26.04.2010
Scientists have combined chemistry and biology research techniques to explain how certain bacteria grow structures on their surfaces that allow them to simultaneously cause illness and protect themselves from the body’s defenses.

The researchers are the first to reproduce a specific component of this natural process in a test tube – an essential step to fully understanding how these structures grow.

With the new method described, these and other researchers now can delve even deeper into the various interactions that must occur for these structures – called lipopolysaccharides – to form, potentially discovering new antibiotic targets along the way.

Lipopolysaccharides are composed primarily of polysaccharides – strings of sugars that are attached to bacterial cell surfaces. They help bacteria hide from the immune system and also serve as identifiers of a given type of bacteria, making them attractive targets for drugs. But before a drug can be designed to inhibit their growth, scientists must first understand how polysaccharides are developed in the first place.

“We were able to answer some of the questions about how components of this growth system do their jobs. This will allow us to more fully characterize lipopolysaccharide biosynthesis in vitro, a process which may shed light on useful targets for developing antibiotic agents,” said Robert Woodward, a graduate student in chemistry at Ohio State University and lead author of the study.

The study is published in the April 25 online edition of the journal Nature Chemical Biology.

The researchers used a harmless strain of Escherichia coli as a model for this work, which would apply to other E. coli strains and similar Gram-negative bacteria, a reference to how their cell walls are structured.

The surface of these bacteria house the lipopolysaccharide, which is a three-part molecular structure embedded into the cell membrane. Two sections of this structure are well understood, but the third, called the O-polysaccharide, has to date been impossible to reproduce.

Two significant challenges have hindered research efforts in this area: The five sugars strung together to compose this section of the molecule are difficult to chemically prepare in the lab, and one of the key enzymes that initiates the structure’s growth process doesn’t easily function in a water-based solution in a test tube.

Ohio State synthetic chemists and biochemists put their heads together to solve these two problems, Woodward said.

To produce the five-sugar chain, the researchers started with a chemically prepared building block containing a single sugar and introduced enzymes that generated a five-sugar unit from that single carbohydrate.

“The first part was done chemically, and in the second part, we used the exact same enzymes that are normally present in a bacterial cell to transform the single sugar into a five-sugar string,” Woodward said.

Once these sugars join to make a five-sugar chain, a specific number of these chains are joined together to fully form the O-polysaccharide. A protein is required to connect those chains – the protein that doesn’t respond well to the test-tube environment.

Early attempts to produce this protein in the lab resulted in clumping structures that did not function. So Woodward and colleagues produced this protein in the presence of what are known as “chaperone” proteins.

“And basically what the chaperones do is help the protein fold into its correct state. We were able to produce the desired enzyme and also were able to verify that it was functional,” Woodward said.

This protein is called Wzy. It is a sugar polymerase, or an enzyme that interacts with the five-sugar chain to begin the process of linking several five-sugar units together.

Getting this far into the process was important, but the researchers also completed one additional step to define yet another protein’s role.

Wzy connected the five-sugar chains, but it did so with no defined limit to the number of five-sugar units involved, a feature that does not match the natural process. On an actual bacterial cell wall, the length of the polysaccharide falls within a relatively narrow range of the number of chains connected.

So the scientists introduced another protein, called Wzz, to the mixture. This protein is known as a “chain length regulator.” With this protein in the mix, the lengths of the resulting polysaccharides were confined to a much more narrow range.

“We were able to replicate the exact polysaccharide biosynthetic pathway in vitro, getting the correct lengths,” Woodward said. “This is important because now you can begin to look at a whole host of other properties in the system.”

The group already started trying to answer one compelling question: whether the two proteins, Wzy and Wzz, have to interact to fully achieve formation of the polysaccharide.

“We’ve shown in some preliminary results that they do interact, but we haven’t determined whether that interaction has any functional relevance,” Woodward said.

With this knowledge in hand, researchers now have access to information about how all three parts of the lipopolysaccharide, the large biomolecule on Gram-negative bacteria cell surfaces, is formed. One thing they already knew is that the entire process takes place on an inner membrane and is then exported to the outer membrane on the cell surface.

Now that scientists can reproduce formation of the lipopolysaccharide, they can more directly characterize the export process – a step in the pathway that serves as another potential antibiotic target, Woodward noted.

This work was supported by the National Institutes of Health, including its Predoctoral Trainee Program, the China Scholarship Council, the National Cancer Institute, the National Science Foundation and the Bill & Melinda Gates Foundation.

Co-authors on the study are Wen Yi, Lei Li, Guohui Zhao, Hironobu Eguchi, Perali Ramu Sridhar, Hongjie Guo, Jing Katherine Song, Edwin Motari, Li Cai, Patrick Kelleher, Xianwei Liu, Weiqing Han, Wenpeng Zhang and Mei Li, all former or current Ohio State graduate students or postdoctoral researchers in biochemistry and chemistry; Yan Ding of Shandong University in China; and Peng George Wang, Ohio Eminent Scholar and professor of biochemistry and chemistry at Ohio State.

Contact: Robert Woodward, (614) 292-8704; woodward.69@osu.edu
Written by Emily Caldwell, (614) 292-8310; caldwell.151@osu.edu

Robert Woodward | EurekAlert!
Further information:
http://www.osu.edu

More articles from Life Sciences:

nachricht Topologische Quantenchemie
21.07.2017 | Max-Planck-Institut für Chemische Physik fester Stoffe

nachricht Topological Quantum Chemistry
21.07.2017 | Max-Planck-Institut für Chemische Physik fester Stoffe

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

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

Im Focus: Manipulating Electron Spins Without Loss of Information

Physicists have developed a new technique that uses electrical voltages to control the electron spin on a chip. The newly-developed method provides protection from spin decay, meaning that the contained information can be maintained and transmitted over comparatively large distances, as has been demonstrated by a team from the University of Basel’s Department of Physics and the Swiss Nanoscience Institute. The results have been published in Physical Review X.

For several years, researchers have been trying to use the spin of an electron to store and transmit information. The spin of each electron is always coupled...

Im Focus: The proton precisely weighted

What is the mass of a proton? Scientists from Germany and Japan successfully did an important step towards the most exact knowledge of this fundamental constant. By means of precision measurements on a single proton, they could improve the precision by a factor of three and also correct the existing value.

To determine the mass of a single proton still more accurate – a group of physicists led by Klaus Blaum and Sven Sturm of the Max Planck Institute for Nuclear...

Im Focus: On the way to a biological alternative

A bacterial enzyme enables reactions that open up alternatives to key industrial chemical processes

The research team of Prof. Dr. Oliver Einsle at the University of Freiburg's Institute of Biochemistry has long been exploring the functioning of nitrogenase....

Im Focus: The 1 trillion tonne iceberg

Larsen C Ice Shelf rift finally breaks through

A one trillion tonne iceberg - one of the biggest ever recorded -- has calved away from the Larsen C Ice Shelf in Antarctica, after a rift in the ice,...

Im Focus: Laser-cooled ions contribute to better understanding of friction

Physics supports biology: Researchers from PTB have developed a model system to investigate friction phenomena with atomic precision

Friction: what you want from car brakes, otherwise rather a nuisance. In any case, it is useful to know as precisely as possible how friction phenomena arise –...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

Closing the Sustainability Circle: Protection of Food with Biobased Materials

21.07.2017 | Event News

»We are bringing Additive Manufacturing to SMEs«

19.07.2017 | Event News

The technology with a feel for feelings

12.07.2017 | Event News

 
Latest News

NASA looks to solar eclipse to help understand Earth's energy system

21.07.2017 | Earth Sciences

Stanford researchers develop a new type of soft, growing robot

21.07.2017 | Power and Electrical Engineering

Vortex photons from electrons in circular motion

21.07.2017 | Physics and Astronomy

VideoLinks
B2B-VideoLinks
More VideoLinks >>>