Hierarchically-structured metalloprotein composite coatings biofabricated from co-existing condensed liquid phases

Franziska Jehle,Elena Macías-Sánchez,Sanja Sviben,Matthew J Harrington,Luca Bertinetti,Peter Fratzl

doi:10.1038/s41467-020-14709-y

Franziska Jehle, Elena Macías-Sánchez + Show 4 more

Open Access

https://doi.org/10.1038/s41467-020-14709-y

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Abstract

Complex hierarchical structure governs emergent properties in biopolymeric materials; yet, the material processing involved remains poorly understood. Here, we investigated the multi-scale structure and composition of the mussel byssus cuticle before, during and after formation to gain insight into the processing of this hard, yet extensible metal cross-linked protein composite. Our findings reveal that the granular substructure crucial to the cuticle’s function as a wear-resistant coating of an extensible polymer fiber is pre-organized in condensed liquid phase secretory vesicles. These are phase-separated into DOPA-rich proto-granules enveloped in a sulfur-rich proto-matrix which fuses during secretion, forming the sub-structure of the cuticle. Metal ions are added subsequently in a site-specific way, with iron contained in the sulfur-rich matrix and vanadium coordinated by DOPA-catechol in the granule. We posit that this hierarchical structure self-organizes via phase separation of specific amphiphilic proteins within secretory vesicles, resulting in a meso-scale structuring that governs cuticle function.

Highlights

Complex hierarchical structure governs emergent properties in biopolymeric materials; yet, the material processing involved remains poorly understood
Previous investigations have identified that the precursor proteins forming the cuticle are stored in secretory vesicles in the mussel foot within a region known as the enzyme or cuticle gland (Fig. 1d)[13,26]
Transmission electron microscopy (TEM) imaging of post-stained thin sections of the cuticle gland of Mytilus edulis revealed that each micron-sized secretory vesicle possesses at least two distinctive regions—a darker outer phase and a lighter, less-stained inner phase (Fig. 2a)

Summary

Introduction

Complex hierarchical structure governs emergent properties in biopolymeric materials; yet, the material processing involved remains poorly understood. We investigated the multiscale structure and composition of the mussel byssus cuticle before, during and after formation to gain insight into the processing of this hard, yet extensible metal cross-linked protein composite. Our findings reveal that the granular substructure crucial to the cuticle’s function as a wear-resistant coating of an extensible polymer fiber is pre-organized in condensed liquid phase secretory vesicles These are phase-separated into DOPA-rich protogranules enveloped in a sulfur-rich proto-matrix which fuses during secretion, forming the sub-structure of the cuticle. Metal ions are added subsequently in a site-specific way, with iron contained in the sulfur-rich matrix and vanadium coordinated by DOPA-catechol in the granule We posit that this hierarchical structure self-organizes via phase separation of specific amphiphilic proteins within secretory vesicles, resulting in a meso-scale structuring that governs cuticle function. By examining the different stages of assembly, we gain important new insights into the formation process and function of this complex biological material, with bearing on the design of technically and biomedically relevant composite materials

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