Influence of nanobody binding on fluorescence emission, mobility, and organization of GFP-tagged proteins.

ABSTRACT: Advanced fluorescence microscopy studies require specific and monovalent molecular labeling with bright and photostable fluorophores. This necessity led to the widespread use of fluorescently labeled nanobodies against commonly employed fluorescent proteins (FPs). However, very little is known how these nanobodies influence their target molecules. Here, we tested commercially available nanobodies and observed clear changes of the fluorescence properties, mobility and organization of green fluorescent protein (GFP) tagged proteins after labeling with the anti-GFP nanobody. Intriguingly, we did not observe any co-diffusion of fluorescently labeled nanobodies with the GFP-labeled proteins. Our results suggest significant binding of the nanobodies to a non-emissive, likely oligomerized, form o

SUBMITTER: Schneider F 

PROVIDER: S-EPMC7753935 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature