In this project, we use a genetically engineered mutant of a bacterial phosphate binding protein to which a coumarin fluorescent probe has been attached. This probe changes its fluorescence by a factor of 5 upon binding of inorganic phosphate, which is tight, specific and fast. This protein is allowed
to diffuse into permeabilised muscle fibres which are then observed in an epi-fluorescence microscope. The release of Pi resulting from the actomyosin ATPase is initiated by the photolytic release of ATP from caged-ATP.
The relevant references can be reached through reference 24 in my publication list.
From left to right
The picture on the left shows Zhen-He He in front of the apparatus. Dr. He is performing all the experiments.
The next picture taken in July 1996 shows the team involved in the application of this technique to the study of smooth muscle contraction. From left to right: Michael Ferenczi, Andrew Somlyo, Avril Somlyo, David Trentham and Zhen-He He. Martin Webb and Martin Brune who developped the probe, and Rod Chillingworth who designed the apparatus are not in the picture.
The next picture shows the computer-controlled rotating stage where the muscle fibres are mounted, allowing rapid changes to the solution surrounding the fibre.
The next picture shows Ger Stienen and Zhen-He He, on the occasion of Dr. Stienen's visit to the laboratory in August 1996 when experiments on rabbit soleus muscle were carried out.
The last picture shows Rod Chillingworth after installation of the sarcomere length measurement system and new motor based on a loudspeaker coil which he designed.
Pictures of the experimental set-up after installation of the sarcomere signal in summer 1997.
page modified 28/11/97 MAF
page modified 24/10/2001 MAF