Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

Critical step traced in anthrax infection

29.07.2005


Pore protein plays active role in toxins’ entry into cells



Scientists at Harvard Medical School (HMS) have revealed details of a key step in the entry of anthrax toxin into human cells. The work, which grew out of an ongoing effort to produce a better anthrax therapeutic, shows that the protective antigen component of the bacterial toxin plays an active role in transferring the other two components of the toxin through the cell membrane. The research, led by R. John Collier, professor of microbiology and molecular genetics at HMS, provides insight into the broader question of how proteins cross cell membranes. The findings appear in the July 29 issue of Science.

An anthrax bacterium secretes three nontoxic proteins that assemble into a toxic complex on the surface of the host cell to set off a chain of events leading to cell toxicity and death. Protective antigen (PA) is one of these proteins, and after binding to the cell, seven copies of it assemble into a specific complex that is capable of forming a pore in a cellular membrane. The pore permits the other two proteins, lethal factor (LF) and edema factor (EF), to enter the cell interior, where the factors interfere with metabolic processes, leading to death of the infected individual.


Details surrounding this process are continuing to be uncovered in Collier’s lab. "Until now, we have not known whether the PA pore serves simply as a passive conduit, or alternatively, plays an active role in shepherding the unfolded LF and EF molecules through," he said. The findings show that it is the latter?the pore takes an active role in protein translocation.

The scientists demonstrated this role by investigating the channel’s chemical make-up. Using a procedure known as cysteine-scanning mutagenesis, they identified the hydrophobic, or "greasy," amino acid phenylalanine in protective antigen’s pore-forming domain. Seven of these amino acids project into the lumen of the pore and form a collection of greasy residues, nicknamed "the phi-clamp" by the scientists. Because the water-filled lumen of the membrane pore is smaller than the folded lethal factor and edema factor, these proteins must first unfold before being actively translocated through the heptameric channel. The clamp appears to work as a chaperone, interacting with the hydrophobic sequences on the two factors as they unfold during translocation. The researchers demonstrated that the phi-clamp was critical to infection by mutating the region and thereby blocking translocation of the toxin proteins.

These recent experimental results extend and explain a 1999 discovery by the Collier lab identifying a set of mutations in protective antigen that prevent translocation, some of which represented a new type of antitoxin that may be useful in anthrax treatment.

In the recent work, Collier and his colleagues found that the phi-clamp composes the main conductance-blocking site for hydrophobic drugs, and it is one of their targets for further investigation. "I believe discovery of the phi-clamp will prove to be one of the high points along the path to understanding how translocation occurs in this system," Collier said.

One of the greatest strengths of the experiment, according to Collier, was the integrative use of technologies applied to the testing procedures. Both cellular systems and model electrophysiological membrane systems were used to test the potency of the anthrax toxin. "We tried to bridge reductionist science with the in vivo situation ?we have to do both to make correlations," he said.

The researchers, who were funded by the National Institutes of Health and the National Science Foundation, will continue to study protein unfolding in translocation during anthrax infection, which may prove to be relevant in other biological systems. "This is only a partial picture," Collier said. "There are still major outstanding questions about the overall process that need to be addressed."

Leah Gourley | EurekAlert!
Further information:
http://www.hms.harvard.edu

More articles from Life Sciences:

nachricht One step closer to reality
20.04.2018 | Max-Planck-Institut für Entwicklungsbiologie

nachricht The dark side of cichlid fish: from cannibal to caregiver
20.04.2018 | Veterinärmedizinische Universität Wien

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

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

Im Focus: Spider silk key to new bone-fixing composite

University of Connecticut researchers have created a biodegradable composite made of silk fibers that can be used to repair broken load-bearing bones without the complications sometimes presented by other materials.

Repairing major load-bearing bones such as those in the leg can be a long and uncomfortable process.

Im Focus: Writing and deleting magnets with lasers

Study published in the journal ACS Applied Materials & Interfaces is the outcome of an international effort that included teams from Dresden and Berlin in Germany, and the US.

Scientists at the Helmholtz-Zentrum Dresden-Rossendorf (HZDR) together with colleagues from the Helmholtz-Zentrum Berlin (HZB) and the University of Virginia...

Im Focus: Gamma-ray flashes from plasma filaments

Novel highly efficient and brilliant gamma-ray source: Based on model calculations, physicists of the Max PIanck Institute for Nuclear Physics in Heidelberg propose a novel method for an efficient high-brilliance gamma-ray source. A giant collimated gamma-ray pulse is generated from the interaction of a dense ultra-relativistic electron beam with a thin solid conductor. Energetic gamma-rays are copiously produced as the electron beam splits into filaments while propagating across the conductor. The resulting gamma-ray energy and flux enable novel experiments in nuclear and fundamental physics.

The typical wavelength of light interacting with an object of the microcosm scales with the size of this object. For atoms, this ranges from visible light to...

Im Focus: Basel researchers succeed in cultivating cartilage from stem cells

Stable joint cartilage can be produced from adult stem cells originating from bone marrow. This is made possible by inducing specific molecular processes occurring during embryonic cartilage formation, as researchers from the University and University Hospital of Basel report in the scientific journal PNAS.

Certain mesenchymal stem/stromal cells from the bone marrow of adults are considered extremely promising for skeletal tissue regeneration. These adult stem...

Im Focus: Like a wedge in a hinge

Researchers lay groundwork to tailor drugs for new targets in cancer therapy

In the fight against cancer, scientists are developing new drugs to hit tumor cells at so far unused weak points. Such a “sore spot” is the protein complex...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

VideoLinks
Industry & Economy
Event News

Invitation to the upcoming "Current Topics in Bioinformatics: Big Data in Genomics and Medicine"

13.04.2018 | Event News

Unique scope of UV LED technologies and applications presented in Berlin: ICULTA-2018

12.04.2018 | Event News

IWOLIA: A conference bringing together German Industrie 4.0 and French Industrie du Futur

09.04.2018 | Event News

 
Latest News

Magnetic nano-imaging on a table top

20.04.2018 | Physics and Astronomy

Start of work for the world's largest electric truck

20.04.2018 | Interdisciplinary Research

Atoms may hum a tune from grand cosmic symphony

20.04.2018 | Physics and Astronomy

VideoLinks
Science & Research
Overview of more VideoLinks >>>