Misaminoacylation of 3,4-dihydroxyphenylalanine (Dopa) molecules to tRNA(Tyr) by endogenous tyrosyl-tRNA synthetase allowed the quantitative replacement tyrosine residues with a yield over 90 % an in vivo residue-specific incorporation strategy, create, for first time, engineered mussel adhesive proteins (MAPs) Escherichia coli very high Dopa content, close that natural MAPs. The Dopa-incorporated MAPs exhibited superior surface adhesion and water resistance ability assistance Dopa-mediated...

Although Dopa-Fe3+ complexation is known to play an important role in mussel adhesion for providing mechanical properties, its function at the plaque/substrate interface, where actual surface occurs, remains unknown, with regard interfacial adhesive proteins (MAPs) type 3 fast variant (fp-3F) and 5 (fp-5). Here, we confirmed of MAPs investigated effects regarding both cohesion. The force measurements using forces apparatus (SFA) analysis showed that intrinsic strong low pH, which similar...

Coacervation of mussel adhesive proteins (MAPs) is proposed as a potential strategy that mussels may use during secretion due to their high concentration density, lack dispersion into seawater, and low interfacial tension. Particularly, coacervations MAPs, foot protein type-3 fast variant (fp-3F) type-5 (fp-5), are important in the initial adhesion process relationship between easy secretion/surface wetting properties coacervate primer-like surface role which directly contact marine surface....

Understanding binding epitopes involved in protein–protein interactions and accurately determining their structure are long-standing goals with broad applicability industry biomedicine. Although various experimental methods for epitope determination exist, these approaches typically low throughput cost-intensive. Computational have potential to accelerate predictions; however, recently developed artificial intelligence (AI)-based frequently fail predict of synthetic domains few natural...

Abstract Background In nature, mussel adhesive proteins (MAPs) show remarkable properties, biocompatibility, and biodegradability. Thus, they have been considered promising biomaterials for various biomedical industrial applications. However, limited production of natural MAPs has hampered their practical Recombinant in bacterial cells could be one alternative to obtain useable amounts MAPs, although additional post-translational modification tyrosine residues into...

Following surgical resection for primary treatment of solid tumors, systemic chemotherapy is commonly used to eliminate residual cancer cells prevent tumor recurrence. However, its clinical outcome often limited due insufficient local accumulation and the toxicity anticancer drugs. Here, we propose a sprayable adhesive nanoparticle (NP)-based drug delivery system using bioengineered mussel protein (MAP) effective locoregional therapy. The MAP NPs could be administered target surfaces in...

We used single-molecule AFM force spectroscopy (AFM-SMFS) in combination with click chemistry to mechanically dissociate anticalin, a non-antibody protein binding scaffold, from its target (CTLA-4), by pulling eight different anchor residues. found that on the anticalin residue 60 or 87 resulted significantly higher rupture forces and decrease koff 2–3 orders of magnitude over range 50–200 pN. Five six internal points gave rise complexes more stable than N- C-terminal points, rupturing at up...

Mussel byssi form a robust underwater adhesive system, anchoring to various surfaces in harsh marine environments. Central byssus is foot protein type 4 (fp-4), junction connecting collagenous threads proteinaceous plaque. This study investigated an anionic plaque-binding domain of fp-4 (fp-4a) and its interactions with cationic proteins (fp-1, fp-5, fp-151 as model substitutes for fp-2) metal ions (Ca2+, Fe3+, V3+). Aggregation, liquid-solid phase transition, was confirmed recombinant fp-4a...

Abstract Mussel adhesive protein (MAP) type 3 (fp-3) is considered one of the key components for mussel adhesion. However, its bulk strength has not been characterized due to availability in limited quantities. In present work, a feasible production (∼47 mg l−1) recombinant fp-3 was achieved, and measured first time; ∼0.57 MPa unmodified form ∼0.94 ∼2.28 3,4-dihydroxy-L-phenylalanine (DOPA)-modified form, having 9.6% yield without with oxidant treatment, respectively. Furthermore, values...

Understanding binding epitopes involved in protein-protein interactions and accurately determining their structure is a long standing goal with broad applicability industry biomedicine. Although various experimental methods for epitope determination exist, these approaches are typically low throughput cost intensive. Computational have potential to accelerate predictions, however, recently developed artificial intelligence (AI)-based frequently fail predict of synthetic domains few natural...

Abstract Catch bonds are a rare class of protein-protein interactions where the bond lifetime increases under an external pulling force. Here, we report how modification anchor geometry generates catch bonding behavior for mechanostable Dockerin G:Cohesin E (DocG:CohE) adhesion complex found on human gut bacteria. Using AFM single-molecule force spectroscopy in combination with bioorthogonal click chemistry, mechanically dissociate using five precisely controlled geometries. When tension is...

Abstract Misaminoacylation of 3,4‐dihydroxyphenylalanine (Dopa) molecules to tRNA Tyr by endogenous tyrosyl‐tRNA synthetase allowed the quantitative replacement tyrosine residues with a yield over 90 % an in vivo residue‐specific incorporation strategy, create, for first time, engineered mussel adhesive proteins (MAPs) Escherichia coli very high Dopa content, close that natural MAPs. The Dopa‐incorporated MAPs exhibited superior surface adhesion and water resistance ability assistance...

Key cellular functions depend on the transduction of extracellular mechanical signals by specialized membrane receptors including adhesion G-protein coupled (aGPCRs). While recently solved structures support aGPCR activation through shedding GAIN domain, molecular mechanisms underpinning receptor mechanosensing remain poorly understood. When probed using single-molecule atomic force spectroscopy and simulations, ADGRG1 dissociated from its tethered agonist at forces significantly higher than...

We investigated the influence of fluorination on unfolding and unbinding reaction pathways a mechanostable protein complex comprising tandem dyad XModule-Dockerin bound to Cohesin. Using single-molecule atomic force spectroscopy, we mapped energy landscapes governing reactions. then used sense codon suppression substitute trifluoroleucine in place canonical leucine globally XMod-Doc. Although TFL substitution thermally destabilized XMod-Doc, it had little effect XMod-Doc:Coh binding affinity...

A spider with mussel: a supramolecular fiber formed by dragline protein was tuned to have underwater adhesion property genetic fusion of mussel foot protein.

Tyrosinase efficiently catalyzes the ortho-hydroxylation of monophenols and oxidation diphenols without any additional cofactors. Although it is significant interest for biosynthesis catechol derivatives, rapid oxidase activity inactivation tyrosinase have hampered its practical utilization as a monophenol monooxygenase. Here, we prepared functional that exhibited distinguished monophenolase/diphenolase ratio (V max mono/ V di = 3.83) enhanced catalytic efficiency against L-tyrosine (k cat...

The opportunistic pathogen Staphylococcus epidermidis utilizes a multidomain surface adhesin protein to bind host components and adhere tissues. While it is known that the interaction between SdrG receptor its fibrinopeptide target (FgB) exceptionally mechanostable (∼2 nN), influence of downstream B domains (B1 B2) unclear. Here, we studied mechanical relationships folded bound FgB. We used engineering, single-molecule force spectroscopy (SMFS) with an atomic microscope (AFM), Monte Carlo...

Abstract Mussel adhesive proteins (MAPs) have great potential as bioglues, particularly in wet conditions. Although vivo residue‐specific incorporation of 3,4‐dihydroxyphenylalanine (Dopa) tyrosine‐auxotrophic Escherichia coli cells allows for production Dopa‐incorporated bioengineered MAPs ( d MAPs), the low yield hinders practical application MAPs. This is due to translational activity a noncanonical amino acid, Dopa, E. cells. Herein, enhance MAPs, we investigated coexpression...

3,4-Dihydroxyphenylalanine (Dopa) is a versatile molecule that enables marine mussels to achieve successful underwater adhesion. However, due its complicated redox chemistry and vulnerability oxidation, controlling surface adhesion cohesion has been challenging issue overcome. Foot protein type 6 (fp-6), thiol-rich interfacial mussel adhesive protein, reported as proteinaceous antioxidant for helps Dopa maintain ability. In this study, we focused on the role of fp-6 in oxidized Dopa. The...