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Optical control of nanobody-mediated protein activity modulation with a photocleavable fluorescent protein.
- Source :
- Analyst; 7/21/2024, Vol. 149 Issue 14, p3865-3870, 6p
- Publication Year :
- 2024
-
Abstract
- Antibodies are crucial in various biological applications due to their specific binding to target molecules, altering protein function and structure. The advent of single-chain antibodies such as nanobodies has paved the way for broader applicability in both research and therapies due to their small size and efficient tissue penetration. Recently, several approaches have been reported to optically control the antigen-binding affinity of nanobodies. Here, we show an alternative strategy for creating photo-activatable nanobodies. By fusing the photocleavable protein PhoCl with the N-terminus of the nanobody (named optoNb60), we successfully demonstrated light-dependent restoration of the antigen-binding ability and the following modulation of the activity of a target protein, the beta-2 adrenergic receptor. Moreover, the activation of optoNb60 was monitored by the fluorescence changes upon photoconversion. The compatibility of the uncaging design with the previously reported optogenetic molecules using nanobodies will contribute to the further optimization of the response capabilities of existing optogenetic tools, thereby expanding their applicability. [ABSTRACT FROM AUTHOR]
- Subjects :
- FLUORESCENT proteins
OPTICAL control
PROTEINS
PROTEIN structure
BETA adrenoceptors
Subjects
Details
- Language :
- English
- ISSN :
- 00032654
- Volume :
- 149
- Issue :
- 14
- Database :
- Complementary Index
- Journal :
- Analyst
- Publication Type :
- Academic Journal
- Accession number :
- 178314139
- Full Text :
- https://doi.org/10.1039/d4an00433g