Biology subjects

Guillem-Marti, J.

Publications and source records attributed to Guillem-Marti, J..

2 recordsLinked to original sources

bioRxiv2024

De Novo Design of Integrin α5β1 Modulating Proteins for Regenerative Medicine

Integrin 5{beta}1 is crucial for cell attachment and migration in development and tissue regeneration, and 5{beta}1 binding proteins could have considerable utility in regenerative medicine and next-generation therapeutics. We use computational protein design to create de novo 5{beta}1-specific modulating miniprotein binders, called NeoNectins, that bind to and stabilize the open state of 5{beta}1. When immobilized onto titanium surfaces and throughout 3D hydrogels, the NeoNectins outperform native fibronectin and RGD peptide in enhancing cell attachment and spreading, and NeoNectin-grafted titanium implants outperformed fibronectin and RGD-grafted implants in animal models in promoting tissue integration and bone growth. NeoNectins should be broadly applicable for tissue engineering and biomedicine. One-Sentence SummaryA de novo-designed fibronectin substitute, NeoNectin, is specific for integrin 5{beta}1 and can be incorporated into biomaterials for regenerative medicine.

bioengineering↗

bioRxiv2024

Multispectral live-cell imaging with uncompromised spatiotemporal resolution

Multispectral imaging is an established method to extend the number of colours usable in fluorescence imaging beyond the typical limit of three or four, but standard approaches are poorly suited to live-cell imaging. We introduce an approach for multispectral imaging in live cells, comprising an iterative spectral unmixing algorithm and eight channel camera-based image acquisition hardware. This enables the accurate unmixing of low signal-to-noise ratio datasets, typical of live-cell imaging, captured at video rates. We use this approach on a commercial spinning disk confocal microscope and a home-built oblique plane light sheet microscope to image 1-7 spectrally distinct fluorophore species simultaneously, using both fluorescent protein fusions and small-molecule dyes. We further use de novo designed protein-binding proteins (minibinders), labelled with organic fluorophores, and use these in combination with our multispectral imaging approach to study the endosomal trafficking of cell-surface receptors at endogenous levels.

biophysics↗