TY - JOUR
T1 - A virus-based single-enzyme nanoreactor
AU - Comellas-Aragones, M.
AU - Engelkamp, H.
AU - Claessen, V.I.
AU - Sommerdijk, N.A.J.M.
AU - Rowan, A.E.
AU - Christianen, P.C.M.
AU - Maan, J.C.
AU - Verduin, B.J.M.
AU - Cornelissen, J.J.L.M.
AU - Nolte, R.J.M.
PY - 2007
Y1 - 2007
N2 - Most enzyme studies are carried out in bulk aqueous solution, at the so-called ensemble level, but more recently studies have appeared in which enzyme activity is measured at the level of a single molecule, revealing previously unseen properties1, 2, 3, 4. To this end, enzymes have been chemically or physically anchored to a surface, which is often disadvantageous because it may lead to denaturation. In a natural environment, enzymes are present in a confined reaction space, which inspired us to develop a generic method to carry out single-enzyme experiments in the restricted spatial environment of a virus capsid. We report here the incorporation of individual horseradish peroxidase enzymes in the inner cavity of a virus, and describe single-molecule studies on their enzymatic behaviour. These show that the virus capsid is permeable for substrate and product and that this permeability can be altered by changing pH.
AB - Most enzyme studies are carried out in bulk aqueous solution, at the so-called ensemble level, but more recently studies have appeared in which enzyme activity is measured at the level of a single molecule, revealing previously unseen properties1, 2, 3, 4. To this end, enzymes have been chemically or physically anchored to a surface, which is often disadvantageous because it may lead to denaturation. In a natural environment, enzymes are present in a confined reaction space, which inspired us to develop a generic method to carry out single-enzyme experiments in the restricted spatial environment of a virus capsid. We report here the incorporation of individual horseradish peroxidase enzymes in the inner cavity of a virus, and describe single-molecule studies on their enzymatic behaviour. These show that the virus capsid is permeable for substrate and product and that this permeability can be altered by changing pH.
KW - fluorescence correlation spectroscopy
KW - tobacco-mosaic-virus
KW - protein
KW - molecule
KW - cage
KW - encapsulation
KW - organization
U2 - 10.1038/nnano.2007.299
DO - 10.1038/nnano.2007.299
M3 - Article
SN - 1748-3387
VL - 2
SP - 635
EP - 639
JO - Nature Nanotechnology
JF - Nature Nanotechnology
IS - 10
ER -