Development of fluorescent biosensors for probing the function of motor proteins.
Biosensors are becoming widely used both in basic research and in screening assays and reagentless sensors with fluorescent reporter groups attached to proteins form one class. This article describes the development of sensors for two small molecules, driven in particular by the need for high sensitivity and time resolution to probe mechanistic aspects of ATP-coupled motor proteins. The biosensors are for the products of the ATPase reaction, ADP and inorganic phosphate. The interplay between the possibilities for design and understanding the mechanism of the signal are discussed. Examples are described of how these sensors have been applied to understanding myosin and helicase motors.
Webb
Martin R
MR
MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London, UK NW7 1AA. mwebb@nimr.mrc.ac.uk
D000244:Adenosine Diphosphate / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D003374:Coumarins / Q000737:chemistry*; D033902:Periplasmic Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D028044:Phosphate-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship
D003374:Coumarins; D033902:Periplasmic Binding Proteins; D028044:Phosphate-Binding Proteins; D010710:Phosphates; C088410:N-(2-(1-maleimidyl)ethyl)-7-(diethylamino)coumarin-3-carboxamide; D000244:Adenosine Diphosphate