biology

+
medicine

The Protein
Switch
A New Technique Improves the Study of Proteins
by Quynh Anh Nguyen
©sxc.hu/Hannah Boettcher

Two models of the FKBP protein used in the experiment.

R
ecent advances made at Stanford University’s
Departments of Chemistry and Chemical and Systems
Biology will now allow scientists to control the function
of a protein more rapidly through the administration of
small molecules. In a paper published in the September 8th issue of
Cell, Dr. Thomas Wandless and his team describe a revolutionary

Photo Credit: http://en.wikipedia.org/wiki/Image:Fkbp-surface-1fkj.png

technique that allows researchers to control the stability of specific
proteins in mammalian cells. This breakthrough has the potential
to make the study of proteins dramatically better, faster, and more
efficient.

Proteins and Their Basic Job Description

Proteins are long strands of amino acids that control
almost every cellular function, including the synthesis and
destruction of tissues, cell signaling, cell cycle control and
regulation, and the immune response. Sections of DNA called
genes contain the information that codes for specific proteins.
This information is first transcribed into a messenger RNA
(mRNA) molecule, which is then translated, or decoded, to
produce the amino acid sequence. These chains of amino
acids then fold into the complex structure of a functional
protein. in the test subject’s system. Therefore, researchers must
wait—sometimes for several days—for the protein to exit

Dr. Thomas Wandless and his the system before continuing onto the next step in their
research protocol. Another disadvantage to this technique
is that it may not be successful in completely inhibiting the
research team have production of a particular protein.

identified a molecule that acts A New On/Off Switch

as a switch to control protein
function.
Traditional Methods of Studying Proteins
Researchers often try to decipher the role a protein
plays by knocking out or knocking down the expression
of the protein product and observing the resulting effects.
Traditionally, this is done by making part of the gene that
codes for a specific protein inactive. Unfortunately, any
protein created prior to inactivation of the gene remains
Wandless and his research team have identified a
molecule that acts as a switch to control protein function.
In their experiment, a destabilizing domain was fused to the
end of a protein of interest by manipulating the DNA that
codes for the protein. When the modified DNA sequence
was translated into a protein, the destabilizing domain
was expressed as an attachment of the protein. Proteins
expressed in this manner were found to be quickly degraded
and completely inactivated. Next, the research team
synthesized a protein dubbed Shield-1 (Shld1) that inhibited
degradation by binding to the destabilizing domain. The
result was that scientists were able to control the presence of

26 stanford scientific

specific proteins more easily and reliably by controlling the
presence of Shld1. In the presence of Shld1, the protein can
be stably expressed, while in the absence of Shld1 the protein
is quickly degraded. Under this new technique, the waiting
time for the protein to degrade is reduced to an average of
four hours.
Proteins expressed in this
manner were found to be
quickly degraded and
completely inactivated.
Prospects for New Findings
This finding has a significant impact on many types of
research. With this technique, scientists can more accurately
study the effects of a protein at a particular instant in a cell’s
life. After employing the method with several proteins,
Wandless reported, “We have not yet seen any cases where
it doesn’t work.” This new method can be used in a variety
of protein settings to aid with research in areas such as
metabolism and immune response, where reactions generally
happen quickly.
The method also provides three great features of flexibility
to experiments. First, Shield-1 has a high bioavailability
and can be administered directly to the cell or the food of
certain test subjects. Second, the technique is reversible:
when Shield-1 is removed, the target protein is degraded
A) Fusion of DNA that encodes for a destabilizing domain (DD) to DNA
that encodes for the protein of interest (POI) results in creation of the
protein attached with a destabilizing domain that causes the protein to
degrade and become inactive. Addition of a ligand to the destabilizing
domain inhibits degradation. (B) The structural formula of the ligand
Shld1. The R in the larger structure corresponds to where the smaller
structure should be attached.
layout design: Jessica Chia-Rong Lee
biology
+
medicine
1 µM Shld1 - + +/-
cell
designation 1
cell
designation 2
cell
designation 3
Pictures of fibroblast cells fused with the destabilizing domain undergoing
treatment with Shld1. Columns stand for type of treatment, rows stand for
cell designation. In the first column (-), the cells were not given Shld1. In the
second column (+), the cells were treated with 1 mM of Shld1. The protein is
activated and changes the appearance of the cell as it is expressed. In the third
column (+/-), Shld1 ligand was first administered for 24 hrs then withdrawn.
Pictures were taken around 48 hrs after withdrawal of Shld1. As shown, the
cell returns back to the phase seen in the first column where the protein is not
expressed.
and inactivated. Finally, the level of protein expression
can be tuned according to the dosage of the administered
Shield-1 ligand. The ease and flexibility with which this
new technology can be applied makes it a useful tool for
studying protein function. In fact, this approach has already
been employed in experiments at several Stanford and non-
Stanford labs using specimens including human cells.
Currently the method is limited only to mammalian
cells, but Wandless’ team continues to study the different
capabilities of this new technology and is currently applying
their research to yeast cells which are important for genetic
studies.. “I‘d be very surprised if we weren’t successful,”
asserts Wandless. S
QUYNH ANH NGUYEN is a freshman majoring in Biological Sciences
with an intended concentration in Molecular and Cell Biology along
with a co-terminal in Bioengineering. In addition to writing for the
magazine, she also enjoys reading scientific journals and papers,
listening to music, and hanging out with her family and friends.
To Learn More:
For more information, visit the Wandless Lab: http://
wandless.stanford.edu/
volume v 27