Oxidation Of Tyrosine Research Articles

Influence of blocking groups on photo-oxidation of tyrosine and derivatives

One-electron oxidation of tyrosine primarily results in the formation of di-tyrosine, which can induce crosslinks leading to protein damage. In this study, we investigated the 3-carboxybenzophenone-sensitized photo-oxidation of Tyr derivatives through time-resolved and steady-state photolysis under anaerobic conditions to analyze the effects of blocking groups.The mechanism for primary and secondary photoreactions in the sensitized photo-oxidation of Tyr derivatives in aqueous solution was presented based on time-resolved analysis and mass spectrometric characterization of photo-oxidation products. Identified di-Tyr products (in addition to those mentioned more often in the literature, such as 3,3′/3,O’) were in some samples presented together with Tyr-CBH adduct (resulting from radical recombination between the tyrosyl radical and CBH•). This publication discusses the possible coulombic effects of interacting ionic species (sensitizer and quencher) on quenching rate constants and the effect of amine groups and steric factors on the distribution of stable products. However, a crucial finding of this work is that the more blocked the Tyr is, the more di-Tyr isomers are formed, suggesting that Tyr residue in proteins may form several forms of di-Tyr cross-links.

Proton-Coupled Electron Transfer upon Oxidation of Tyrosine in a De Novo Protein: Analysis of Proton Acceptor Candidates.

Redox-active residues, such as tyrosine and tryptophan, play important roles in a wide range of biological processes. The α3Y de novo protein, which is composed of three α helices and a tyrosine residue Y32, provides a platform for investigating the redox properties of tyrosine in a well-defined protein environment. Herein, the proton-coupled electron transfer (PCET) reaction that occurs upon oxidation of tyrosine in this model protein by a ruthenium photosensitizer is studied by using a vibronically nonadiabatic PCET theory that includes hydrogen tunneling and excited vibronic states. The input quantities to the analytical nonadiabatic rate constant expression, such as the diabatic proton potential energy curves and associated proton vibrational wave functions, reorganization energy, and proton donor-acceptor distribution functions, are obtained from density functional theory calculations on model systems and molecular dynamics simulations of the solvated α3Y protein. Two possible proton acceptors, namely, water or a glutamate residue in the protein scaffold, are explored. The PCET rate constant is greater when glutamate is the proton acceptor, mainly due to the more favorable driving force and shorter equilibrium proton donor-acceptor distance, although contributions from excited vibronic states mitigate these effects. Nevertheless, water could be the dominant proton acceptor if its equilibrium constant associated with hydrogen bond formation is significantly greater than that for glutamate. Although these calculations do not definitively identify the proton acceptor for this PCET reaction, they elucidate the conditions under which each proton acceptor can be favored. These insights have implications for tyrosine-based PCET in a wide variety of biochemical processes.

Voltammetric detection of Neuropeptide Y using a modified sawhorse waveform

The hormone Neuropeptide Y (NPY) plays critical roles in feeding, satiety, obesity, and weight control. However, its complex peptide structure has hindered the development of fast and biocompatible detection methods. Previous studies utilizing electrochemical techniques with carbon fiber microelectrodes (CFMEs) have targeted the oxidation of amino acid residues like tyrosine to measure peptides. Here, we employ the modified sawhorse waveform (MSW) to enable voltammetric identification of NPY through tyrosine oxidation. Use of MSW improves NPY detection sensitivity and selectivity by reducing interference from catecholamines like dopamine, serotonin, and others compared to the traditional triangle waveform. The technique utilizes a holding potential of −0.2 V and a switching potential of 1.2 V that effectively etches and renews the CFME surface to simultaneously detect NPY and other monoamines with a sensitivity of 5.8 ± 0.94 nA/µM (n = 5). Furthermore, we observed adsorption-controlled, subsecond NPY measurements with CFMEs and MSW. The effective identification of exogenously applied NPY in biological fluids demonstrates the feasibility of this methodology for in vivo and ex vivo studies. These results highlight the potential of MSW voltammetry to enable fast, biocompatible NPY quantification to further elucidate its physiological roles.Graphical

Class Ib Ribonucleotide Reductases: Activation of a Peroxido-MnIIMnIII to Generate a Reactive Oxo-MnIIIMnIV Oxidant.

In the postulated catalytic cycle of class Ib Mn2 ribonucleotide reductases (RNRs), a MnII2 core is suggested to react with superoxide (O2·-) to generate peroxido-MnIIMnIII and oxo-MnIIIMnIV entities prior to proton-coupled electron transfer (PCET) oxidation of tyrosine. There is limited experimental support for this mechanism. We demonstrate that [MnII2(BPMP)(OAc)2](ClO4) (1, HBPMP = 2,6-bis[(bis(2 pyridylmethyl)amino)methyl]-4-methylphenol) was converted to peroxido-MnIIMnIII (2) in the presence of superoxide anion that converted to (μ-O)(μ-OH)MnIIIMnIV (3) via the addition of an H+-donor (p-TsOH) or (μ-O)2MnIIIMnIV (4) upon warming to room temperature. The physical properties of 3 and 4 were probed using UV-vis, EPR, X-ray absorption, and IR spectroscopies and mass spectrometry. Compounds 3 and 4 were capable of phenol oxidation to yield a phenoxyl radical via a concerted PCET oxidation, supporting the proposed mechanism of tyrosyl radical cofactor generation in RNRs. The synthetic models demonstrate that the postulated O2/Mn2/tyrosine activation mechanism in class Ib Mn2 RNRs is plausible and provides spectral insights into intermediates currently elusive in the native enzyme.

Iron-Mediated Electrolysis for Selective Oxidative Decarboxylation of C-Terminal Acids Using Metal-Binding Residue

With the increasing concern of the environmental impact of solid-phase peptide synthesis (SPPS), semi-recombinant processes, in which recombinantly expressed peptides are selectively modified, are of growing interest. The strategies employed for these modifications must overcome the challenges associated with recombinant peptides: selectivity across many functional groups and solubility of the full-length peptide. Electrochemistry is a viable solution to these challenges as it is a mild and tunable process which can operate in both organic and aqueous systems. Herein, a model tetramer with a redox-active residue, tyrosine, is selectively decarboxylated at the C-terminus in an iron-mediated electrolysis. Chemoselectivity is achieved by exploiting the metal-binding property of a C-terminal glutamic acid residue. In the presence of the iron mediator, the tyrosine oxidation pathway is suppressed while the oxidative decarboxylation pathway is favored. Mechanistic controls demonstrate that the glutamic acid, iron, and applied current are all necessary to impart the desired chemoselectivity.

Site-Selective Tyrosine Reaction for Antibody-Cell Conjugation and Targeted Immunotherapy.

Targeted immunotherapies capitalize on the exceptional binding capabilities of antibodies to stimulate a host response that effectuates long-lived tumor destruction. One example is the conjugation of immunoglobulins (IgGs) to immune effector cells, which equips the cells with the ability to recognize and accurately kill malignant cells through a process called antibody-dependent cellular cytotoxicity (ADCC). In this study, a chemoenzymatic reaction is developed that specifically functionalizes a single tyrosine (Tyr, Y) residue, Y296, in the Fc domain of therapeutic IgGs. A one-pot reaction that combines the tyrosinase-catalyzed oxidation of tyrosine to o-quinone with a subsequent [3+2] photoaddition with vinyl ether is employed. This reaction installs fluorescent molecules or bioorthogonal groups at Y296 of IgGs or the C-terminal Y-tag of an engineered nanobody. The Tyr-specific reaction is utilized in constructing monofunctionalized antibody-drug conjugates (ADCs) and antibody/nanobody-conjugated effector cells, such as natural killer cells or macrophages. These results demonstrate the potential of site-selective antibody reactions for enhancing targeted cancer immunotherapy.

Avoiding one-electron oxidation of tyrosine by DOPA

Avoiding one-electron oxidation of tyrosine by DOPA

Tyrosine Amino Acid as a Foulant for the Heterogeneous Anion Exchange Membrane.

The features of organic fouling have been revealed for highly basic anion exchange membranes during prolonged electrodialysis in solutions containing the aromatic amino acid tyrosine. With increased operation time when using MA-41 heterogeneous membranes in tyrosine solution, an increase in hydrophobicity and roughness characteristics of the material surface is detected. A reduction in tyrosine flux through the membrane occurs which is caused by its pores plugging and deposition of the amino acid at the membrane surface induced by tyrosine adsorption and local supersaturation of the solution in the membrane phase. The long-term contact of the anion exchange membrane with a solution of tyrosine leads to some structural changes in the anion exchange material. An accumulation of the studied amino acid with phenolic fragment and tyrosine oxidation products (DOPA, DOPA-quinone) is found and confirmed by IR- and UV-spectroscopy techniques. The organic fouling is accompanied by an increase in density and a decrease in moisture content of the studied membrane. A comparative analysis of the chemical and electrochemical cleaning results for fouled samples of the MA-41 membrane demonstrates a partial restoration of the material transport characteristics using electrochemical cleaning in the intensive current mode of electrodialysis. The best efficiency of regeneration is reached when carrying out chemical cleaning with a solution of hydrochloric acid, providing almost complete restoration of the membrane characteristics.

Untapped Potential: Real-Time Measurements of Opioid Exocytosis at Single Cells.

The endogenous opioid system is commonly targeted in pain treatment, but the fundamental nature of neuropeptide release remains poorly understood due to a lack of methods for direct detection of specific opioid neuropeptides in situ. These peptides are concentrated in, and released from, large dense-core vesicles in chromaffin cells. Although catecholamine release from these neuroendocrine cells is well characterized, the direct quantification of opioid peptide exocytosis events has not previously been achieved. In this work, a planar carbon-fiber microelectrode served as a "postsynaptic" sensor for probing catecholamine and neuropeptide release dynamics via amperometric monitoring. A constant potential of 500 mV was employed for quantification of catecholamine release, and a higher potential of 1000 mV was used to drive oxidation of tyrosine, the N-terminal amino acid in the opioid neuropeptides released from chromaffin cells. By discriminating the results collected at the two potentials, the data reveal unique kinetics for these two neurochemical classes at the single-vesicle level. The amplitude of the peptidergic signals decreased with repeat stimulation, as the halfwidth of these signals simultaneously increased. By contrast, the amplitude of catecholamine release events increased with repeat stimulation, but the halfwidth of each event did not vary. The chromogranin dense core was identified as an important mechanistic handle by which separate classes of transmitter can be kinetically modulated when released from the same population of vesicles. Overall, the data provide unprecedented insight into key differences between catecholamine and opioid neuropeptide release from isolated chromaffin cells.

Mechanistic Insights into Tyrosine Oxidation Revealed by Fast-Scan Cyclic Voltammetry

A precise understanding of opioid dynamics in the brain is lacking, largely because few analytical tools are available that are sufficiently sensitive and selective for directly monitoring these important neurochemicals in situ. Tyrosine is a key part of the opioid neuropeptides, and its redox activity can be exploited in their detection. However, a solid understanding of tyrosine electrochemistry on carbon electrodes is first required. It is known that, upon oxidation, tyrosine quickly undergoes chemical transformation to products that foul the electrode, compromising the electrochemical information. This issue can be avoided by using fast-scan cyclic voltammetry (FSCV) which enables collection of well-defined voltammograms on a millisecond timescale before appreciable accumulation of byproducts. In this work, the tyrosine oxidation reaction is characterized and electrode fouling is assessed when tyrosine is incorporated into short synthetic peptides containing other amino acids that are part of the opioid neuropeptides: Gly, Phe, Met, Leu, and Arg. Voltammetric data were obtained in phosphate buffered solution on carbon microelectrodes at scan rates up to 1 kV/s. The results demonstrate the dependence of the peak oxidation potential on solution pH, and identify possible reaction pathways and byproducts that foul the electrode. Peak oxidation potential, peak shape, and ultimately, sensitivity to tyrosine are dependent on the peptide sequence. NMR spectroscopy is used to relate the proximity of proton acceptor/donor functional groups to the electrochemical results in the context of proton-coupled electron transfer (PCET) theory. These fundamental studies are important for the electrochemical detection of important neuropeptides in the complex in vivo environment. Furthermore, a better understanding of the tunability of PCET reaction rates in electrochemical systems has broad implications for technology in a range of areas including artificial photosynthesis, solar energy conversion, and energy storage.

A New Thiolate-Bound Dimanganese Cluster as a Structural and Functional Model for Class Ib Ribonucleotide Reductases.

In class Ib ribonucleotide reductases (RNRs) a dimanganese(II) cluster activates superoxide (O2 ⋅- ) rather than dioxygen (O2 ), to access a high valent MnIII -O2 -MnIV species, responsible for the oxidation of tyrosine to tyrosyl radical. In a biomimetic approach, we report the synthesis of a thiolate-bound dimanganese complex [MnII 2 (BPMT)(OAc)2 ](ClO)4 (BPMT=(2,6-bis{[bis(2-pyridylmethyl)amino]methyl}-4-methylthiophenolate) (1) and its reaction with O2 ⋅- to form a [(BPMT)MnO2 Mn]2+ complex 2. Resonance Raman investigation revealed the presence of an O-O bond in 2, while EPR analysis displayed a 16-line St =1/2 signal at g=2 typically associated with a MnIII MnIV core, as detected in class Ib RNRs. Unlike all other previously reported Mn-O2 -Mn complexes, generated by O2 ⋅- activation at Mn2 centers, 2 proved to be a capable electrophilic oxidant in aldehyde deformylation and phenol oxidation reactions, rendering it one of the best structural and functional models for class Ib RNRs.

3D-printed sensor decorated with nanomaterials by CO2 laser ablation and electrochemical treatment for non-enzymatic tyrosine detection

Thecombination of CO2 laser ablation and electrochemical surface treatments is demonstratedto improve the electrochemical performance of carbon black/polylactic acid (CB/PLA) 3D-printed electrodes through the growth of flower-like Na2O nanostructures on their surface. Scanning electron microscopy images revealed that the combination of treatments ablated the electrode's polymeric layer, exposing a porous surface where Na2O flower-like nanostructures were formed. The electrochemical performance of the fabricated electrodes was measured by the reversibility of the ferri/ferrocyanide redox couple presenting a significantly improved performance compared withelectrodes treated by only one of the steps. Electrodes treated by the combined method also showed a better electrochemical response for tyrosine oxidation. These electrodes were used as a non-enzymatic tyrosine sensor for quantification in human urine samples. Two fortified urine samples were analyzed, and the recovery values were 106 and 109%. The LOD and LOQ for tyrosine determination were 0.25 and 0.83 μmol L-1, respectively, demonstrating that the proposed devices are suitable sensors for analyses of biological samples, even at low analyte concentrations.

Avoiding One-Electron Oxidation of Biomolecules by 3,4-Dihydroxy-L-Phenylalanine (DOPA)†.

It has been proposed that 3,4-dihydroxy-L-phenylalanine (DOPA) has antioxidant properties, and thus, the objective of this work was to evaluate the effect of adding DOPA during the photosensitized oxidation of tyrosine (Tyr), tryptophan (Trp), histidine (His), 2'-deoxyguanosine 5'-monophosphate (dGMP) and 2'-deoxyadenosine 5'-monophosphate (dAMP). It was observed that, upon pterin-photosensitized degradation of a given biomolecule in acidic aqueous solutions, the rate of the biomolecule consumption decreases due to the presence of DOPA. Although DOPA deactivates the excited states of pterin (Ptr), biomolecules do as well, being the bimolecular quenching constants in the diffusional control limit, indicating that DOPA antioxidant mechanism is not a simple deactivation of Ptr excited states. Laser flash photolysis experiments provide evidence of the formation of DOPA radical (DOPA(-H)• , λMAX 310 nm), which is formed in a timescale longer than Ptr triplet excited state (3 Ptr*) lifetime, ruling out its formation in a reaction between DOPA and 3 Ptr*. The experimental results presented in this work indicate that the observed decrease on the rate of each biomolecule consumption due to the presence of DOPA is through a second one-electron transfer reaction from DOPA to the biomolecule radicals.

In-situ synthesis of melanin in tumor with engineered probiotics for hyperbaric oxygen-synergized photothermal immunotherapy

Anaerobic bacteria and facultative anaerobe are usually used as delivery carriers of antitumor drugs because of their tumor hypoxia targeting and antitumor immune activation abilities, but suffer from dose-dependent systemic side effects. Here, engineered probiotics Nissle 1917 (EcN-T) were constructed for in-situ synthesis of melanin in tumor region for enhanced photothermal therapy (PTT) combining with immunotherapy. By means of genetic engineering, EcN-T that could highly express tyrosinase under near-infrared (NIR) light irradiation were constructed. Due to their intrinsic tumor hypoxia targeting, EcN-T could accumulate and rapidly proliferate in tumor region after intravenous injection. Tyrosinase expression for the following melanin biosynthesis within tumor could be initiated upon NIR exposure. Hyperbaric oxygen treatment facilitated the oxidation of tyrosine to promote the generation of melanin in situ, giving robust photothermal performance to induce immunogenic tumor cell death. Assited by immunostimulatory responses of EcN, maturation of dendritic cells and priming of cytotoxic T cells were largely promoted in tumor tissue, resulting in significant inhibition of tumor growth and recurrence. This bacteria-mediated combined therapeutic strategy showed high efficacy and good safety.

A Novel Approach to Vaccine Development: Concomitant Pathogen Inactivation and Host Immune Stimulation by Peroxynitrite.

The design of efficient vaccines for long-term protective immunity against pathogens represents an objective of utmost public health priority. In general, live attenuated vaccines are considered to be more effective than inactivated pathogens, yet potentially more reactogenic. Accordingly, inactivation protocols which do not compromise the pathogen’s ability to elicit protective immunity are highly beneficial. One of the sentinel mechanisms of the host innate immune system relies on the production of reactive nitrogen intermediates (RNI), which efficiently inactivate pathogens. Peroxynitrite (PN) is a prevalent RNI, assembled spontaneously upon the interaction of nitric oxide (NO) with superoxide. PN exerts its bactericidal effect by via the efficient oxidation of a broad range of biological molecules. Furthermore, the interaction of PN with proteins results in structural/chemical modifications, such as the oxidation of tryptophan, tyrosine, and cysteine residues, as well as the formation of carbonyl, dityrosine, and nitrotyrosine (NT). In addition to their role in innate immunity, these PN-mediated modifications of pathogen components may also augment the antigenicity of pathogen peptides and proteins, hence contributing to specific humoral responses. In the study reported here, a novel approach for vaccine development, consisting of pathogen inactivation by PN, combined with increased immunity of NT-containing peptides, is implemented as a proof-of-concept for vaccination against the intracellular pathogen Francisella tularensis (F. tularensis). In vivo experiments in a murine model of tularemia confirm that PN-inactivated F. tularensis formulations may rapidly stimulate innate and adaptive immune cells, conferring efficient protection against a lethal challenge, superior to that elicited by bacteria inactivated by the widely used formalin treatment.

Sweat analysis with a wearable sensing platform based on laser-induced graphene.

The scientific community has shown increasing interest in laser scribing for the direct fabrication of conductive graphene-based tracks on different substrates. This can enable novel routes for the noninvasive analysis of biofluids (such as sweat or other noninvasive matrices), whose results can provide the rapid evaluation of a person's health status. Here, we present a wearable sensing platform based on laser induced graphene (LIG) porous electrodes scribed on a flexible polyimide sheet, which samples sweat through a paper sampler. The device is fully laser manufactured and features a two layer design with LIG-based vertical interconnect accesses. A detailed characterization of the LIG electrodes including pore size, surface groups, surface area in comparison to electroactive surface area, and the reduction behavior of different LIG types was performed. The bare LIG electrodes can detect the electrochemical oxidation of both uric acid and tyrosine. Further modification of the surface of the LIG working electrode with an indoaniline derivative [4-((4-aminophenyl)imino)-2,6-dimethoxycyclohexa-2,5-dien-1-one] enables the voltammetric measurement of pH with an almost ideal sensitivity and without interference from other analytes. Finally, electrochemical impedance spectroscopy was used to measure the concentrations of ions through the analysis of the sweat impedance. The device was successfully tested in a real case scenario, worn on the skin during a sports session. In vitro tests proved the non-cytotoxic effect of the device on the A549 cell line.

Sporicidal Action of Pulsed Radiation of Hot Plasma.

In the study, we used a Pilimin IR-10 spark discharge generator as a source of pulsed radiation of hot plasma; a corona discharge generator - as a source of cold plasma; a DKB-9 low-pressure mercury lamp - as a source of continuous radiation of UV band, wavelength of 253.7 nm. The samples were processed in Petri dishes, 40 mm in diameter, their volume being 4 and 10 cm3. The study used an L-tyrosine solution in distilled water (the concentration: 160 mg/L), a suspension of bacteria and spores of micromycetes (its concentration being ~106 cells per 1 ml). Tyrosine conversion products were identified spectrophotometrically before and after treatment. The biocidal and sporicidal effects were assessed by counting CFU (colony-forming units) after seeding incubation at 27-37°C. The oxidation of tyrosine by HO2• radicals was found to be impossible. Under 2 the action of nitrogen compounds, nitration proceeds with 3-nitrotyrosine formation. The nitration reaction is slow, taking about 100 h. A possible nitration mechanism is through the formation of the nitronium ion NO2+ in an acidic medium.The biocidal effect of hot plasma radiation turned out to be weaker than that of UV radiation of a DKB-9 lamp. This is due to the difference in their emission spectrum. The sporicidal effect of hot plasma radiation was more pronounced: a 10-fold decrease in the number of CFU was observed at radiation doses of 200-280 J. Under the action of UV radiation, at the same doses, the decrease in the number of CFU was from 3 to ~30%. The sporicidal effect of hot plasma radiation is due to the decay of a long-living …ONOOH/ONOO-… complex with the formation of a nitric oxide and a nitronium ion in an acidic medium. The study showed the viability of spores under the action of pulsed radiation of hot plasma to decrease. While the light radiation of a UV lamp, under studied conditions, slightly penetrates the protective coating of a spore. The sporicidal effect of hot plasma radiation is due to the decay of a long-living …ONOOH/ONOO-… complex with the formation of a nitric oxide and a nitronium ion in an acidic medium. Nitration plays a decisive role in the sporicidal action of the hot plasma radiation of a spark discharge. The principle of the sporicidal effect of gas-discharge plasma radiation can be used to develop disinfecting devices.

De novo protein design of photochemical reaction centers

Natural photosynthetic protein complexes capture sunlight to power the energetic catalysis that supports life on Earth. Yet these natural protein structures carry an evolutionary legacy of complexity and fragility that encumbers protein reengineering efforts and obfuscates the underlying design rules for light-driven charge separation. De novo development of a simplified photosynthetic reaction center protein can clarify practical engineering principles needed to build new enzymes for efficient solar-to-fuel energy conversion. Here, we report the rational design, X-ray crystal structure, and electron transfer activity of a multi-cofactor protein that incorporates essential elements of photosynthetic reaction centers. This highly stable, modular artificial protein framework can be reconstituted in vitro with interchangeable redox centers for nanometer-scale photochemical charge separation. Transient absorption spectroscopy demonstrates Photosystem II-like tyrosine and metal cluster oxidation, and we measure charge separation lifetimes exceeding 100 ms, ideal for light-activated catalysis. This de novo-designed reaction center builds upon engineering guidelines established for charge separation in earlier synthetic photochemical triads and modified natural proteins, and it shows how synthetic biology may lead to a new generation of genetically encoded, light-powered catalysts for solar fuel production.

Proton-Coupled Electron-Transfer Reduction of Dioxygen: The Importance of Precursor Complex Formation between Electron Donor and Proton Donor.

The proton-coupled electron transfer (PCET) reaction has drawn extensive attention for its widespread occurrence in chemistry, biology, and materials science. The mechanistic studies via model systems such as tyrosine and phenol oxidation have gradually deepened the understanding of PCET reactions, which was widely accepted and applied to bond activation and transformation. However, direct PCET activation of nonpolar bonds such as the C-H bond, O2, and N2 has yet to be explored. Herein, we report that the interaction between electron donor and proton donor could overcome the barrier of direct O2 activation via a concerted electron-proton transfer mechanism. This work provides a new strategy for developing direct PCET activation of nonpolar bonds.

Stable and direct coating of fibronectin-derived Leu-Asp-Val peptide on ePTFE using one-pot tyrosine oxidation for endothelial cell adhesion

Expanded polytetrafluoroethylene (ePTFE) is widely used in clinical applications, such as in the manufacture of blood-contacting implantable devices, owing to its flexibility, biostability, and non-adhesiveness. Modification with peptides is an effective strategy to further improve the ePTFE function. However, the chemical stability of PTFE makes it difficult to modify with peptides. In this study, we reported a simple method for the dense and stable coating of biofunctional peptides on the ePTFE surface through the anchor sequence, Tyr-Lys-Tyr-Lys-Tyr-Lys (YK3). A peptide (YK3-LDV) incorporating the YK3 anchor and a ligand sequence for α4β1 integrin, Leu-Asp-Val (LDV), was successfully coated on ePTFE grafts through one-pot oxidation. The peptide layer constructed via YK3-LDV coating on ePTFE was stable and resistant to extensive washing by aqueous solutions of highly concentrated salts and surfactants. YK3-LDV coating promoted the in vitro adhesion of endothelial cells to ePTFE. Furthermore, YK3-LDV coating accelerated the in vivo formation of neointima-like tissue in a rat model with an ePTFE patch implanted into the carotid artery.