Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/156082
Title: Liquid–liquid phase separation of the green mussel adhesive protein Pvfp-5 is regulated by the post-translated Dopa amino acid
Authors: Deepankumar, Kanagavel
Guo, Qi
Mohanram, Harini
Lim, Jessica
Mu, Yuguang
Pervushin, Konstantin
Yu, Jing
Miserez, Ali
Keywords: Engineering::Materials
Issue Date: 2021
Source: Deepankumar, K., Guo, Q., Mohanram, H., Lim, J., Mu, Y., Pervushin, K., Yu, J. & Miserez, A. (2021). Liquid–liquid phase separation of the green mussel adhesive protein Pvfp-5 is regulated by the post-translated Dopa amino acid. Advanced Materials. https://dx.doi.org/10.1002/adma.202103828
Project: MOE 2019-T3-1-012
NRF-NRFF11-2019-0004
Journal: Advanced Materials 
Abstract: The underwater adhesive prowess of aquatic mussels has been largely attributed to the presence of the post-translationally modified amino acid 3,4-dihydroxylphenylalanine (Dopa) in mussel foot proteins (MFPs) that make up their adhesive threads. In parallel, it has been suggested that during thread fabrication, MFPs form intermediate fluidic phases such as liquid crystals or coacervates during a Liquid-Liquid Phase Separation (LLPS) process. Here, we show that Dopa plays another central role during mussel fiber formation, by enabling LLPS of Pvfp-5, a main MFP of the Asian green mussel Perna viridis. Using residue-specific substitution of Tyrosine (Tyr) for Dopa during recombinant expression, we show that Dopa-substituted Pvfp-5 exhibits LLPS under seawater-like conditions, whereas the Tyr-only version forms insoluble aggregates. Combining quantum chemistry calculations and solution NMR studies, we find that a transient H-bonding network requiring the two hydroxyl groups of Dopa is critical to enable LLPS in Dopa-mutated Pvfp-5. Overall, our study suggests that Dopa plays an important role in regulating LLPS of MFPs, which may be critical to concentrate the adhesive proteins at the plaque/substrate interface and therefore produce a more robust adhesive. Our findings also provide molecular-level lessons to guide biomanufacturing of protein-based materials such as bioadhesives and load-bearing fibers.
URI: https://hdl.handle.net/10356/156082
ISSN: 0935-9648
DOI: 10.1002/adma.202103828
Schools: School of Materials Science and Engineering 
School of Biological Sciences 
Research Centres: Center for Sustainable Materials
Rights: This is the peer reviewed version of the following article: Deepankumar, K., Guo, Q., Mohanram, H., Lim, J., Mu, Y., Pervushin, K., Yu, J. & Miserez, A. (2021). Liquid–liquid phase separation of the green mussel adhesive protein Pvfp-5 is regulated by the post-translated Dopa amino acid. Advanced Materials, which has been published in final form at https://doi.org/10.1002/adma.202103828. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:MSE Journal Articles
SBS Journal Articles

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