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@ARTICLE{Rosenkranz:4647,
author = {Rosenkranz, T. and Katranidis, A. and Atta, D. and
Enderlein, J. and Gregor, I. and Grzelakowski, M. and
Rigler, P. and Meier, W. and Fitter, J.},
title = {{O}bserving {P}roteins as {S}ingle {M}olecules
{E}ncapsulated in {S}urface-{T}ethered {P}olymeric
{N}anocontainers},
journal = {ChemBioChem},
volume = {10},
issn = {1439-4227},
address = {Weinheim},
publisher = {Wiley-VCH},
reportid = {PreJuSER-4647},
pages = {702 - 709},
year = {2009},
note = {Jurgen Groll (SusTech GmbH $\&$ Co KG, Darmstadt) is
greatly acknowledged for providing us with a protocol for
cover slide coatings. We thank Iris v. d. Hocht for
providing Atto655-labeled DOPE. T.R. acknowledges financial
support by the International Helmholtz Research School on
Biophysics and Soft Matter ("Bio-Soft'). J.F thanks G. Buldt
(Forschungszentrum Julich) for continuous and sustainable
support in his institute.},
abstract = {Immobilizing biomolecules provides the advantage of
observing them individually for extended time periods, which
is impossible to accomplish for freely diffusing molecules
in solution. In order to immobilize individual protein
molecules, we encapsulated them in polymeric vesicles made
of amphiphilic triblock copolymers and tethered the vesicles
to a cover slide surface. A major goal of this study is to
investigate polymeric vesicles with respect to their
suitability for protein-folding studies. The fact that
polymeric vesicles possess an extreme stability under
various chemical conditions is supported by our observation
that harsh unfolding conditions do not perturb the
structural integrity of the vesicles. Moreover,
polymerosomes prove to be permeable to GdnHCl and, thereby,
ideally suited for unfolding and refolding studies with
encapsulated proteins. We demonstrate this with encapsulated
phosphoglycerate kinase, which was fluorescently labeled
with Atto655, a dye that exhibits pronounced photoinduced
electron transfer (PET) to a nearby tryptophan residue in
the native state. Under unfolding conditions, PET was
reduced, and we monitored alternating unfolding and
refolding conditions for individual encapsulated proteins.},
keywords = {Fluorescent Dyes: metabolism / Fungal Proteins: chemistry /
Fungal Proteins: metabolism / Hydrophobic and Hydrophilic
Interactions / Immobilized Proteins: chemistry / Immobilized
Proteins: metabolism / Liposomes: chemistry / Nanoparticles:
chemistry / Photochemical Processes / Polymers: chemistry /
Protein Denaturation / Protein Folding / Protein
Renaturation / Saccharomyces cerevisiae / Surface Properties
/ Fluorescent Dyes (NLM Chemicals) / Fungal Proteins (NLM
Chemicals) / Immobilized Proteins (NLM Chemicals) /
Liposomes (NLM Chemicals) / Polymers (NLM Chemicals) / J
(WoSType)},
cin = {ISB-2},
ddc = {540},
cid = {I:(DE-Juel1)ISB-2-20090406},
pnm = {Programm Biosoft},
pid = {G:(DE-Juel1)FUEK443},
shelfmark = {Biochemistry $\&$ Molecular Biology / Chemistry, Medicinal},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:19191249},
UT = {WOS:000264168000015},
doi = {10.1002/cbic.200800739},
url = {https://juser.fz-juelich.de/record/4647},
}