title: Directed supramolecular surface assembly of SNAP-tag fusion proteins.
abstract: Supramolecular assembly of proteins on surfaces and vesicles was investigated by site-selective incorporation of a supramolecular guest element on proteins. Fluorescent proteins were site-selectively labeled with bisadamantane by SNAP-tag technology. The assembly of the bisadamantane functionalized SNAP-fusion proteins on cyclodextrin-coated surfaces yielded stable monolayers. The binding of the fusion proteins is specific and occurs with an affinity in the order of 10(6) M(-1) as determined by surface plasmon resonance. Reversible micropatterns of the fusion proteins on micropatterned cyclodextrin surfaces were visualized by using fluorescence microscopy. Furthermore, the guest-functionalized proteins could be assembled out of solution specifically onto the surface of cyclodextrin vesicles. The SNAP-tag labeling of proteins thus allows for assembly of modified proteins through a host-guest interaction on different surfaces. This provides a new strategy in fabricating protein patterns on surfaces and takes advantage of the high labeling efficiency of the SNAP-tag with designed supramolecular elements.
authors.affiliation: Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands.
authors.forename: Dana A
authors.initials: DA
authors.lastname: Uhlenheuer
authors.forename: Dorothee
authors.initials: D
authors.lastname: Wasserberg
authors.forename: Christian
authors.initials: C
authors.lastname: Haase
authors.forename: Hoang D
authors.initials: HD
authors.lastname: Nguyen
authors.forename: Jan Hendrik
authors.initials: JH
authors.lastname: Schenkel
authors.forename: Jurriaan
authors.initials: J
authors.lastname: Huskens
authors.forename: Bart Jan
authors.initials: BJ
authors.lastname: Ravoo
authors.forename: Pascal
authors.initials: P
authors.lastname: Jonkheijm
authors.forename: Luc
authors.initials: L
authors.lastname: Brunsveld
mesh_terms: D003505:Cyclodextrins / Q000737:chemistry*; D007103:Immobilization; D046911:Macromolecular Substances / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D053835:Unilamellar Liposomes / Q000737:chemistry*
chemical_list: D003505:Cyclodextrins; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D053835:Unilamellar Liposomes