Triggering enzymatic activity with force.
Integrating single molecule force spectroscopy with fluorescence-based techniques allows the manipulation of an enzyme with a periodic stretching and relaxation protocol while simultaneously monitoring its catalytic activity. After releasing the stretching force we observe a higher probability for enzymatic activity at a time of 1.7 s. A detailed theoretical analysis reveals that the relaxation from the force-induced enzyme conformation to the observed active conformation follows a cascade reaction with several steps and a free energy difference of at least 8 k(B)T. Our study clearly points out the direct influence of force on enzymatic activity and opens up a new way to study and manipulate (bio)catalytic reactions at the single molecule level.
Gumpp
Hermann
H
Lehrstuhl für Angewandte Physik and Center for Nanoscience, Center for Integrated Protein Science Munich, LMU München, D-80799 München, Germany.
Puchner
Elias M
EM
Zimmermann
Julia L
JL
Gerland
Ulrich
U
Gaub
Hermann E
HE
Blank
Kerstin
K
D002384:Catalysis; D005452:Fluoresceins / Q000737:chemistry; D005453:Fluorescence; D005656:Fungal Proteins; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D036103:Nanotechnology
D005452:Fluoresceins; D005656:Fungal Proteins; C024098:6-carboxyfluorescein; C093616:6-carboxyfluorescein diacetate; C045132:4-carboxyfluorescein; C084577:5-carboxyfluorescein diacetate; D008049:Lipase; C431349:lipase B, Candida antarctica