Featured Research

from universities, journals, and other organizations

Proteins shine a brighter light on cellular processes: Cyan fluorescent protein will make cellular imaging more sensitive

Date:
March 20, 2012
Source:
European Synchrotron Radiation Facility
Summary:
Scientists have designed a molecule which, in living cells, emits turquoise light three times brighter than possible until recently. This improves the sensitivity of cellular imaging, a technique where biological processes inside a living organism are imaged at high resolution.

This image is an artistically inspired visualization of the three-dimensional X-ray structure of the Cyan Fluorescence Protein mTurquoise2.
Credit: Nature Communications/von Stetten/Royant/Goedhart

Scientists have designed a molecule which emits turquoise light more efficiently than ever seen before in living cells. This improves the sensitivity of cellular imaging, a technique for high-resolution imaging of biological processes inside a living cell.

The results have been published in Nature Communications on 20 March 2012.

The lead author of the publication is Antoine Royant from the Institut de Biologie Structurale/CNRS-CEA-Universitι Joseph Fourier in Grenoble. The team also comprised scientists from the Universities of Amsterdam and Oxford, and the European Synchrotron Radiation Facility (ESRF) in Grenoble.

Cyan fluorescent proteins (CFPs) are very popular in cell biology where they are used to make processes visible inside a living cell, like in a movie. These processes include the changes in the shape of large biological molecules. Since the early 1990s, fluorescent proteins have become one of the most important tools used in the biosciences and have helped the observation of previously invisible processes such as the development of nerve cells in the brain or the spread of cancer cells. The 2008 Nobel Prize in Chemistry crowned their discovery and rapid development.

CFPs allow mapping of many processes in living cells when they can be attached to a protein involved in an interaction or a conformational change. The CFP inside the cell is localised by illuminating the cell with blue light which makes the fluorescent protein emit light of a characteristic colour, which is cyan for CFPs. The emission of light from the CFP reveals the target of observation, the protein to which it is attached. However, these molecules have long suffered from a weak fluorescence level, converting merely 36% of the incoming blue into cyan light.

To achieve higher brightness, and with it improved sensitivity of fluorescent imaging, the scientists based in France, led by Antoine Royant, teamed up with colleagues from the Netherlands and the United Kingdom.

First, using highly brilliant X-ray beams at the ESRF, the teams from Grenoble and Oxford uncovered subtle details of how CFPs store incoming energy and retransmit it as fluorescent light: they produced tiny crystals of many different improved CFPs and resolved their molecular structures. These structures revealed a subtle process near the so-called chromophore, the light-emitting complex inside the CFPs, whose fluorescence efficiency could be modulated by the environment. "We could understand the function of individual atoms within CFPs and pinpoint the part of the molecule that needed to be modified to increase the fluorescence yield" says David von Stetten from the ESRF.

In parallel to this work, the Amsterdam team led by Theodorus Gadella used an innovative screening technique to study hundreds of modified CFP molecules, measuring their fluorescence lifetimes under the microscope to identify which had improved properties.

The result of this rational design is a new CFP, called mTurquoise2. By combining structural and cellular biology efforts, the researchers managed to show that mTurquoise2 has a fluorescence efficiency of 93%, unmatched for this type of protein.

The new molecule will allow life scientists to study protein-protein interactions in living cells with unprecedented sensitivity. High sensitivity matters in processes where only a few proteins are involved and signals are weak, and in fast reactions where the time available for accumulating fluorescent light is short.

"With the new protein, many studies can now be performed with levels of accuracy and detail that were impossible yesterday. Moreover, thanks to this novel approach taking into account the structural dynamics of the protein, scientists now hope to design improved fluorescent proteins emitting light of different colours for use in other applications," concludes Antoine Royant.


Story Source:

The above story is based on materials provided by European Synchrotron Radiation Facility. Note: Materials may be edited for content and length.


Journal Reference:

  1. Joachim Goedhart, David von Stetten, Marjolaine Noirclerc-Savoye, Mickaλl Lelimousin, Linda Joosen, Mark A. Hink, Laura van Weeren, Theodorus W.J. Gadella, Antoine Royant. Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%. Nature Communications, 2012; 3: 751 DOI: 10.1038/ncomms1738

Cite This Page:

European Synchrotron Radiation Facility. "Proteins shine a brighter light on cellular processes: Cyan fluorescent protein will make cellular imaging more sensitive." ScienceDaily. ScienceDaily, 20 March 2012. <www.sciencedaily.com/releases/2012/03/120320141938.htm>.
European Synchrotron Radiation Facility. (2012, March 20). Proteins shine a brighter light on cellular processes: Cyan fluorescent protein will make cellular imaging more sensitive. ScienceDaily. Retrieved July 31, 2014 from www.sciencedaily.com/releases/2012/03/120320141938.htm
European Synchrotron Radiation Facility. "Proteins shine a brighter light on cellular processes: Cyan fluorescent protein will make cellular imaging more sensitive." ScienceDaily. www.sciencedaily.com/releases/2012/03/120320141938.htm (accessed July 31, 2014).

Share This




More Plants & Animals News

Thursday, July 31, 2014

Featured Research

from universities, journals, and other organizations


Featured Videos

from AP, Reuters, AFP, and other news services

Raw: Thousands Flocking to German Crop Circle

Raw: Thousands Flocking to German Crop Circle

AP (July 30, 2014) — Thousands of people are trekking to a Bavarian farmer's field to check out a mysterious set of crop circles. (July 30) Video provided by AP
Powered by NewsLook.com
Concern Grows Over Worsening Ebola Crisis

Concern Grows Over Worsening Ebola Crisis

AFP (July 30, 2014) — Pan-African airline ASKY has suspended all flights to and from the capitals of Liberia and Sierra Leone amid the worsening Ebola health crisis, which has so far caused 672 deaths in Guinea, Liberia and Sierra Leone. Duration: 00:43 Video provided by AFP
Powered by NewsLook.com
At Least 20 Chikungunya Cases in New Jersey

At Least 20 Chikungunya Cases in New Jersey

AP (July 30, 2014) — At least 20 New Jersey residents have tested positive for chikungunya, a mosquito-borne virus that has spread through the Caribbean. (July 30) Video provided by AP
Powered by NewsLook.com
Xtreme Eating: Your Daily Caloric Intake All On One Plate

Xtreme Eating: Your Daily Caloric Intake All On One Plate

Newsy (July 30, 2014) — The Center for Science in the Public Interest released its 2014 list of single meals with whopping calorie counts. Video provided by Newsy
Powered by NewsLook.com

Search ScienceDaily

Number of stories in archives: 140,361

Find with keyword(s):
 
Enter a keyword or phrase to search ScienceDaily for related topics and research stories.

Save/Print:
Share:  

Breaking News:
from the past week

In Other News

... from NewsDaily.com

Science News

Health News

    Environment News

    Technology News



      Save/Print:
      Share:  

      Free Subscriptions


      Get the latest science news with ScienceDaily's free email newsletters, updated daily and weekly. Or view hourly updated newsfeeds in your RSS reader:

      Get Social & Mobile


      Keep up to date with the latest news from ScienceDaily via social networks and mobile apps:

      Have Feedback?


      Tell us what you think of ScienceDaily -- we welcome both positive and negative comments. Have any problems using the site? Questions?
      Mobile iPhone Android Web
      Follow Facebook Twitter Google+
      Subscribe RSS Feeds Email Newsletters
      Latest Headlines Health & Medicine Mind & Brain Space & Time Matter & Energy Computers & Math Plants & Animals Earth & Climate Fossils & Ruins