Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

Chemists Develop More Efficient Protein Labeling

07.02.2012
North Carolina State University researchers have created specially engineered mammalian cells to provide a new “chemical handle” which will enable researchers to label proteins of interest more efficiently, without disrupting the normal function of the proteins themselves or the cells in which they are found.

Protein labeling is used by researchers in a variety of fields to help them understand how these important molecules affect the normal functioning of cells. Currently, proteins are labeled for study simply by fusing them to other fluorescent proteins, which allows researchers to use microscopy to track their movements through a cell. This approach has several drawbacks, however, not least being that the fluorescent proteins are often large enough to affect the function of the protein of interest.

Dr. Alex Deiters, associate professor of chemistry, along with colleague Dr. Jason Chin of the Laboratory of Molecular Biology at the Medical Research Council in Cambridge, U.K., have developed a way to attach a fluorophore – a fluorescent molecule about 20 times smaller than the fluorescent proteins currently in use – to a protein that is expressed in a mammalian cell.

Deiters and Chin developed a special 21st amino acid that they added to cells that were specially engineered to incorporate this amino acid into the protein they wanted to study (there are normally only 20 amino acids). This 21st amino acid has a “chemical handle” that only reacts with a specifically designed fluorophore, but not any cellular components. According to Deiters, “The reaction between the modified protein and the fluorophore is extremely fast, high yielding, and generates a stable link between both reaction partners. This novel methodology enables future cell biological studies that were previously not possible.”

The research appears in the Feb. 5 issue of Nature Chemistry.

“We found that our approach gave us a higher yield of labeled proteins and that the binding reaction was 50 times faster than with current methods,” Deiters says. “Additionally, it took less reagent to complete the reaction, so overall we have a faster, more efficient method for protein labeling, and less chance of interfering with the normal function of the proteins and cells being studied.”

The research was funded by the National Institutes of Health and the National Science Foundation. The Department of Chemistry is part of NC State’s College of Physical and Mathematical Sciences.

Note to editors: Abstract of the paper follows

“Genetically encoded norbornene directs site-specific cellular protein labelling via a

rapid bioorthogonal reaction”

Authors: Alexander Deiters, Jessica Torres-Kolbus, Chungjung Chou, North Carolina State University; Jason W. Chin, Kathrin Lang, Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge, UK

Published: Feb. 5, 2012 in Nature Chemistry

Abstract:
The site-specific incorporation of bioorthogonal groups via the expansion of genetic code provides a powerful general strategy for site-specifically labelling proteins with any probe. However, the slow reactivity of the bioorthogonal functional groups that can be encoded genetically limits the utility of this strategy. We demonstrate the genetic encoding of a norbornene amino acid using the pyrrolysyl transfer RNA synthetase/tRNACUA pair in Escherichia coli and mammalian cells. We developed a series of tetrazine-based probes that exhibit ‘turn-on’ fluorescence on their rapid reaction with norbornenes. We demonstrate that the labelling of an encoded norbornene is specific with respect to the entire soluble E. coli proteome and thousands of times faster than established encodable bioorthogonal reactions. We show explicitly the advantages of this approach over state-of-the-art bioorthogonal reactions for protein labelling in vitro and on mammalian cells, and so demonstrate the first rapid bioorthogonal site-specific labelling of a protein on the mammalian cell surface.

Tracey Peake | Newswise Science News
Further information:
http://www.ncsu.edu

More articles from Life Sciences:

nachricht Toward a 'smart' patch that automatically delivers insulin when needed
18.01.2017 | American Chemical Society

nachricht 127 at one blow...
18.01.2017 | Stiftung Zoologisches Forschungsmuseum Alexander Koenig, Leibniz-Institut für Biodiversität der Tiere

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

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

Im Focus: How gut bacteria can make us ill

HZI researchers decipher infection mechanisms of Yersinia and immune responses of the host

Yersiniae cause severe intestinal infections. Studies using Yersinia pseudotuberculosis as a model organism aim to elucidate the infection mechanisms of these...

Im Focus: Interfacial Superconductivity: Magnetic and superconducting order revealed simultaneously

Researchers from the University of Hamburg in Germany, in collaboration with colleagues from the University of Aarhus in Denmark, have synthesized a new superconducting material by growing a few layers of an antiferromagnetic transition-metal chalcogenide on a bismuth-based topological insulator, both being non-superconducting materials.

While superconductivity and magnetism are generally believed to be mutually exclusive, surprisingly, in this new material, superconducting correlations...

Im Focus: Studying fundamental particles in materials

Laser-driving of semimetals allows creating novel quasiparticle states within condensed matter systems and switching between different states on ultrafast time scales

Studying properties of fundamental particles in condensed matter systems is a promising approach to quantum field theory. Quasiparticles offer the opportunity...

Im Focus: Designing Architecture with Solar Building Envelopes

Among the general public, solar thermal energy is currently associated with dark blue, rectangular collectors on building roofs. Technologies are needed for aesthetically high quality architecture which offer the architect more room for manoeuvre when it comes to low- and plus-energy buildings. With the “ArKol” project, researchers at Fraunhofer ISE together with partners are currently developing two façade collectors for solar thermal energy generation, which permit a high degree of design flexibility: a strip collector for opaque façade sections and a solar thermal blind for transparent sections. The current state of the two developments will be presented at the BAU 2017 trade fair.

As part of the “ArKol – development of architecturally highly integrated façade collectors with heat pipes” project, Fraunhofer ISE together with its partners...

Im Focus: How to inflate a hardened concrete shell with a weight of 80 t

At TU Wien, an alternative for resource intensive formwork for the construction of concrete domes was developed. It is now used in a test dome for the Austrian Federal Railways Infrastructure (ÖBB Infrastruktur).

Concrete shells are efficient structures, but not very resource efficient. The formwork for the construction of concrete domes alone requires a high amount of...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

12V, 48V, high-voltage – trends in E/E automotive architecture

10.01.2017 | Event News

2nd Conference on Non-Textual Information on 10 and 11 May 2017 in Hannover

09.01.2017 | Event News

Nothing will happen without batteries making it happen!

05.01.2017 | Event News

 
Latest News

A big nano boost for solar cells

18.01.2017 | Power and Electrical Engineering

Glass's off-kilter harmonies

18.01.2017 | Materials Sciences

Toward a 'smart' patch that automatically delivers insulin when needed

18.01.2017 | Life Sciences

VideoLinks
B2B-VideoLinks
More VideoLinks >>>