Single molecule reaction analysis of P-types ATPase using electrochemical fast-scanning atomic force microscopy

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Ion pumps transport specific cationic ion across the biomembrane against a concentration gradient through the energy obtained from ATP hydrolysis. Recent progresses in crystallographic study of sarcoplasmic reticulum Ca^<2+>-ATPase (SERCA), which is one of the best-characterized P-type ATPases, allow us to discuss the transport mechanisms involving dynamical structural changes of P-type ATPases.In this study, we constructed a novel method to analyze molecular reaction of P-type ATPase at a single molecular level and real time scale using a newly developed fast-scanning atomic force microscopy. Using the system, we succeeded to observe single molecule of SERCA and plasma membrane sodium pump act as a pumping machine by direct detection of their repeatedly up-and-down conformational changes.