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Photoactivatable Hsp47: A tool to control and regulate collagen secretion & assembly

2018, Khan, Essak, Sankaran, Shrikrishnan, Paez, Julieta, Muth, Christina, Han, Mitchell, del Campo, Aránzazu

Hsp47 is a chaperone protein with a fundamental role in the folding, stability and intracellular transport of procollagen triple helices. A light-responsive Hsp47 recombinant protein, engineered to control in situ the production and assembly of cellular collagen is here demonstrated. This novel light-driven tool enables unprecedented fundamental studies of collagen biosynthesis and associated diseases.

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A photoactivatable α5β1-specific integrin ligand

2018, Vakkeel, Roshna, Aleeza, Farrukh, del Campo, Aranzazu

In order to study how dynamic changes of α5β1 integrin engagement affect cellular behaviour, photoactivatable derivatives of α5β1 specific ligands are presented in this article. The presence of the photoremovable protecting group (PRPG) introduced at a relevant position for integrin recognition, temporally inhibits ligand bioactivity. Light exposure at cell-compatible dose efficiently cleaves the PRPG and restores functionality. Selective cell response (attachment, spreading, migration) to the activated ligand on the surface is achieved upon controlled exposure. Spatial and temporal control of the cellular response is demonstrated, including the possibility to in situ activation. Photoactivatable integrin-selective ligands in model microenvironments will allow the study of cellular behavior in response to changes in the activation of individual integrins as consequence of dynamic variations of matrix composition.

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Possibilities and Limitations of Photoactivatable Cytochalasin D for the Spatiotemporal Regulation of Actin Dynamics

2020, Nair, Roshna V., Zhao, Shifang, Terriac, Emmanuel, Lautenschläger, Franziska, Hetmanski, Joseph H.R., Caswell, Patrick T., del Campo, Aranzazu

The study of the actin cytoskeleton and related cellular processes requires tools to specifically interfere with actin dynamics in living cell cultures, ideally with spatiotemporal control and compatible with real time imaging. A phototriggerable derivative of the actin disruptor Cytochalasin D (CytoD) is described and tested here. It includes a nitroveratryloxycarbonyl (Nvoc) photoremovable protecting group (PPG) at the hydroxyl group at C7 of CytoD. The attachment of the PPG renders Nvoc-CytoD temporarily inactive, and enables light-dosed delivery of the active drug CytoD to living cells. This article presents the full structural and physicochemical characterization, the toxicity analysis. It is complemented with biological tests to show the time scales (seconds) and spatial resolution (cellular level) achievable with a UV source in a regular microscopy setup