Cross-Linking and Bundling of Self-Assembled Protein-Based Polymer Fibrils via Heterodimeric Coiled Coils

Natalia E. Domeradzka, Marc W.T. Werten, Frits A. De Wolf, Renko De Vries*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

6 Citations (Scopus)

Abstract

Previously, we developed triblock protein polymers that form fibrillar hydrogels at low protein polymer concentrations (denoted C2-SH 48-C2). We here demonstrate that the structure of these hydrogels can be tuned via heterodimeric coiled coils that cross-link and bundle the self-assembled protein polymer fibrils. We fused well-characterized, 47 amino acids-long heterodimeric coiled coil "linkers" (DA or DB) to the C-terminus of the triblock polymer. The resulting C2-SH 48-C2-DA and C2-SH 48-C2-DB polymers, were successfully produced as secreted proteins in Pichia pastoris, with titers of purified protein in the order of g L-1 of clarified broth. Atomic force microscopy showed that fibrils formed by either C2-SH 48-C2-DA or C2-SH 48-C2-DB alone already displayed extensive bundling, apparently as a result of homotypic (DA/DA and DB/DB) interactions. For fibrils prepared from protein polymers having no linkers, plus a small fraction of polymers containing either DA or DB linkers, no cross-linking and bundling was observed. At these same low concentrations of linkers, fibrils containing both the DA and the DB linkers did show cross-linking and bundling as a consequence of heterodimer formation. This work shows that we can control the extent of bundling and cross-linking of supramolecular fibrils by varying the density of heterodimerizing coiled coils in the fibrils, which is promising for the further development of materials that mimic the extracellular matrix.
Original languageEnglish
Pages (from-to)3893-3901
JournalBiomacromolecules
Volume17
Issue number12
DOIs
Publication statusPublished - 2016

Fingerprint

Dive into the research topics of 'Cross-Linking and Bundling of Self-Assembled Protein-Based Polymer Fibrils via Heterodimeric Coiled Coils'. Together they form a unique fingerprint.

Cite this