Following a protein's travel inside cells is key to improving patient monitoring, drug development

Apr 23, 2014
Virginia Tech chemical engineer Chang Lu, and his graduate student Zhenning Cao, right, are authors of the journal article about a unique process that could help in drug development. Credit: Virginia Tech

In science, "simple and accessible detection methods that can rapidly screen a large cell population with the resolution of a single cell inside that population has been seriously lacking," said Virginia Tech chemical engineer Chang Lu.

In the Royal Society of Chemistry journal Chemical Science, Lu has announced that he and his coworkers have developed a novel technique that detects the subcellular location of a . The significance of this method is that the technique will allow the scientific and technological communities a simple and improved method for studying effectiveness of therapies for disease, including cancer.

If a protein is not located in the right subcellular compartment of a cell, "the result can be diseases ranging from metabolic disorders to cancers," Lu explained. "Modulation of inside a cell is practiced as an important therapeutic approach for cancer treatment. The subcellular location of a can also serve as a useful read-out for high-content screening of cancer drugs."

In the human body, proteins move between distinct compartments inside cells, including the , the nucleus, and other membrane-enclosed areas. This movement can be a prerequisite for proteins to carry out their intended functions. These functions might include gene transcription and other molecular regulations.

In the publication, Lu and his colleagues, Zhenning Cao, a graduate student of the School of Biomedical Engineering and Sciences, Shuo Geng, a postdoctoral researcher, and Liwu Li, professor of biological sciences, all of Virginia Tech, described the shortcomings of current evaluations of protein movement.

One such technique is fluorescence microscopy. According to Lu, it can only analyze a limited number of cells. Data collected by a second existing assay called subcellular fractionation only reflects the average properties of the cell populations "without revealing the heterogeneity that is often present among seemingly identical cells," they stated.

Lu' team had made some progress in screening in the past using an electroporation-based technique, but it did not allow the examination of native proteins and primary cells isolated from animals and from patients.

Their new work uses a method that "incorporates selective chemical release of cytosolic proteins with a standard procedure for fluorescent labeling of the protein to detect the subcellular location of a native protein," Lu said. This simple and unique tweak to the conventional cell staining process allowed them to accurately define the subcellular location of the protein by measuring the amount of the residual protein after release. Using a flow cytometer, the speed of such measurement could reach 10,000 to 100,000 per second.

A key ingredient to their process is the use of saponin, a class of amphipathic glycosides. It dissolves cholesterol and permeates the plasma membrane to allow protein release. "Gentle treatment by saponin shows minimal effects on the state of the cell," Lu added.

Explore further: Efficient analysis of small quantity of cells improves chances to understand disease

More information: Chem. Sci., 2014, DOI: 10.1039/C4SC00578C, http://pubs.rsc.org/en/content/articlelanding/2014/sc/c4sc00578c#!divAbstract

add to favorites email to friend print save as pdf

Related Stories

Watching the production of new proteins in live cells

Aug 26, 2013

Researchers at Columbia University, in collaboration with biologists in Baylor College of Medicine, have made a significant step in understanding and imaging protein synthesis, pinpointing exactly where and ...

Recommended for you

Molecules that came in handy for first life on Earth

Nov 24, 2014

For the first time, chemists have successfully produced amino acid-like molecules that all have the same 'handedness', from simple building blocks and in a single test tube. Could this be how life started. ...

User comments : 0

Please sign in to add a comment. Registration is free, and takes less than a minute. Read more

Click here to reset your password.
Sign in to get notified via email when new comments are made.