Subcellular visualization and quantification of cyanotoxins microcystin or anabaenopeptin synthesis in bloom-forming cyanobacteria shows a distinct compartmentation

ABSTRACT: In this study time-lapse experiments have been performed using pulsed feeding of non-natural amino acids (non-AAs) in order to observe the build up or decline of azide- or alkyne-modified (clickable) microcystins (MCs) in M. aeruginosa. Clickable MCs have been subsequently labeled via Copper-catalyzed Azide-Alkyne Cycloaddition (CuAAC) with ALEXA Fluor 488 (A488) and imaged using confocal microscopy analysis and analysed using Huygens 23.10 software. To confirm the specific labeling of clickable peptides, monoclonal primary antibody for MC was used for comparison with chemoselective labeling of clickable MCs in M. aeruginosa. Additionally polyclonal antibody against Rubisco large subunit form I and form II was used as a control of the performance of the immunolabeling reaction. For this purpose, a double labeling protocol for chemo-selective labeling of clickable MCs (A555) and immunolabeling of proteins (A488) was established.

SUBMITTER: Rubén Morón Asensio 

PROVIDER: S-BIAD2489 | bioimages |

REPOSITORIES: bioimages