Protection of the indole nucleus of tryptophan in solid-phase peptide synthesis with a dipeptide that can be cleaved rapidly at physiological pH

The synthesis of a new derivative of tryptophan Fmoc-Trp(Boc-Sar-Sar)-OH is described. Fmoc-Trp(Boc-Sar-Sar)-OH is introduced into peptides by solid-phase peptide synthesis and during treatment with TFA at the end of the synthesis, the Boc group is cleaved and the peptide is obtained with the indole nucleus modified with the sarcosinyl-sarcosinyl (Sar-Sar) moiety. This modification will introduce a cationic charge that improves the solubility of the peptide during HPLC purification. The Sar-Sar moiety is cleaved upon exposure to physiological pH. The Boc-Sar-Sar group might, therefore, also find use in the design of pro-drugs of indole-containing compounds.     

Simultaneous determination of ascorbic acid,dopamine and uric acid based on tryptophan functionalized graphene

A new type of tryptophan-functionalized graphene nanocomposite (Trp-GR) was synthesized by utilizing a facile ultrasonic method via π–π conjugate action between graphene (GR) and tryptophan (Trp) molecule. The material as prepared had well dispersivity in water and better conductivity than pure GR. The surface morphology of Trp-GR was characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Raman spectroscopy. The electrochemical behaviors of ascorbic acid (AA), dopamine (DA), and uric acid (UA) were investigated by cyclic voltammetry (CV) on the surface of Trp-GR. The separation of the oxidation peak potentials for AA–DA, DA–UA and UA–AA was about 182 mV, 125 mV and 307 mV, which allowed simultaneously determining AA, DA, and UA. Differential pulse voltammetery (DPV) was used for the determination of AA, DA, and UA in their mixture. Under optimum conditions, the linear response ranges for the determination of AA, DA, and UA were 0.2–12.9 mM, 0.5–110 μM, and 10–1000 μM, with the detection limits (S/N = 3) of 10.09 μM, 0.29 μM and 1.24 μM, respectively. Furthermore, the modified electrode was investigated for real sample analysis.     

A comprehensive conformational analysis of tryptophan,its ionic and dimeric forms

Tryptophan is an essential amino acid, and understanding the conformational preferences of monomer and dimer is a subject of outstanding relevance in biological systems. An exhaustive first principles investigation of tryptophan ( W ) and its ionized counterparts cations (WC) , anions (WA) , and zwitterions (WZ) has been carried out. A comprehensive and systematic study of tryptophan dimer (WD) conformations resulted in about 62 distinct minima on the potential energy surface. The hydrogen bonds and a variety of noncovalent interactions such as OH‐π, NH‐π, CH‐π, CH‐O, and π‐π interactions stabilized different forms of tryptophan and its dimers. Over all in monomeric conformers which have NH‐O, hydrogen bonds showed higher stability than other conformers. A cursory analysis reveal that the most stable dimers stabilized by hydrogen bonding interactions while the less stable dimers showed aromatic side chain interactions. Protein Data Bank analysis of tryptophan dimers reveals that at a larger distance greater than 5 Å, T‐shaped orientations (CH‐π interactions) are more prevalent, while stacked orientations (π‐π interactions) are predominant at a smaller distance. © 2013 Wiley Periodicals, Inc.     

Microchip electrophoresis with chemiluminescence detection for assaying ascorbic acid and amino acids in single cells

A method based on microchip electrophoresis (MCE) with chemiluminescence (CL) detection was developed for the determination of ascorbic acid (AA) and amino acids including tryptophan (Trp), glycine (Gly) and alanine (Ala) present in single cells. Cell injection, loading, lysing, electrophoretic separation and CL detection were integrated onto a simple cross microfluidic chip. A single cell was loaded in the cross intersection by electrophoretic means through applying a set of potentials at the reservoirs. The docked cell was lysed rapidly under a direct electric field. The intracellular contents were MCE separated within 130 s. CL detection was based on the enhancing effects of AA and amino acids on the CL reaction of luminol with K₃[Fe(CN)₆]. Rat hepatocytes were prepared and analyzed as the test cellular model. The average intracellular contents of AA, Trp, Gly and Ala in single rat hepatocytes were found to be 38.3, 5.15, 3.78 and 3.84 fmol (n = 12), respectively.     

Electrochemical behavior and voltammetric determination of tryptophan based on 4-aminobenzoic acid polymer film modified glassy carbon electrode

Herein, a novel electrochemical method was developed for the determination of tryptophan based on the poly(4-aminobenzoic acid) film modified glassy carbon electrode (GCE). The electrochemical behaviors of tryptophan at the modified electrode were investigated. It was found that the oxidation peak current of tryptophan at the modified GCE was greatly improved compared with that at the bare GCE. The effects of supporting electrolyte, pH value, scan rate, accumulation potential and time were examined. The oxidation peak current of tryptophan was proportional to its concentration over the range from 1.0 × 10⁻⁶ to 1.0 × 10⁻⁴ mol L⁻¹. The limit of detection was evaluated to be 2.0 × 10⁻⁷ mol L⁻¹. The proposed method was sensitive and simple. It was successfully employed to determine tryptophan in pharmaceutical samples.     

荧光探针法研究色氨酸二肽与DNA的相互作用

以溴化乙锭(EB)为荧光探针,对色氨酸二肽(Trp-Trp)与小牛胸腺DNA的作用机制进行了研究,考查了酸度、离子强度及磷酸根离子对Trp-rrp与DNA相互作用的影响;结果表明,在ph 7.2的模拟体液条件下,Tro-Trp可与DNA发生相互作用;向DNA-EB体系中加入Trp-Trp时荧光逐渐减弱,这说明Trp-Trp 能与DNA发生作用.通过ScatehaId方程进一步分析发现,Trp-Trp对DNA-EB的影响表现为混合模式:一方面,Trp-Trp能中和DNA上的磷氧负离子,导致DNA链收缩,从而影响DNA的构象,使嵌入的EB从DNA中部分游离出来,荧光减弱;另一方面,Trp-Trp与EB也存在静电位点竞争.     

Fluorescence-quenching of Tryptophan Ion-18-crown-6 Complex——A Different Observation from Its Neutral Counterpart

It is well-known that primary amines form inclusive complexes with crown ethers. This kind of noncova-lently inclusive complexes,exemplified tryptophan complex with 18-crown-6,was studied by combination of two experimental methods,fluorimetry and mass spectrometry,with theoretical calculation. Fluorescence intensity of tryptophan in the absence and the presence of 18-crown-6 in aqueous HAc-NaAc buffer solution at pH=3.73 has been studied by fluorometry. The fluorescence intensity decreases significantly wit...     

Synthesis of 5-[(2-hydroxynaphthalen-1-yl)diazenyl]isophthalic acid and its application to electrocatalytic oxidation and determination of adrenaline,paracetamol, and tryptophan

In this study,Au nanoparticles/poly 5-[(2-hydroxynaphthalen-l-yl)diazenyl]isophthalic acid film modified glassy carbon electrode(AuNPs/poly(NDI)/GCE) has shown excellent electrocatalytic activity toward the oxidation of adrenaline(ADR),paracetamol(PAC),and tryptophan(Trp).The bare glassy carbon electrode(GCE) fails to separate the oxidation peak potentials of these molecules,while the poly(NDI) film modified electrode can resolve them.Electrochemical impedance spectroscopy(EIS)indicates that the charge transfer resistance of the bare electrode decreases as 5-[(2-hydroxynaphthalen-l-yl)diazenyl]isophthalic acid is electropolymerized on the bare electrode.Furthermore,EIS exhibits enhancement of electron transfer kinetics between analytes and the electrode after electrodeposition of Au nanoparticles.Differential pulse voltammetry results show that the electrocatalytic current increases linearly in the ranges of 0.01-680.0 μmol L~1 for ADR,0.05-498.0 μmol L~1 for PAC,and 3.0-632.0 μmol L~1 for Trp;with detection limits(S/N = 3) of 0.009 μmol L~1,0.005 μmol L~1,and 0.09 μmol L~1 for ADR,PAC,and Trp,respectively.The proposed method has been successfully applied for simultaneous determination of ADR,PAC,and Trp in biological samples.     

Destabilization of artificial biomembrane induced by the penetration of tryptophan

The effect of tryptophan on the membrane stability was studied by using three artificial biological membranes including liposome, Langmuir monolayer and solid supported bilayer lipid membrane (s-BLM) as models. All the results indicate that the penetration of tryptophan can destabilize different artificial biological membranes. The diameter of liposome and the leakage of calcein from liposome increased with the increase of tryptophan concentration because the penetration of tryptophan was beneficial for dehydrating the polar head groups of lipids and the formation of fusion intermediates. π-A isotherms of lecithin on the subphase of tryptophan solution further confirm that tryptophan can penetrate into lipid monolayer and reduce the stability of lipid monolayer. When the concentration of tryptophan increased from 0 to 2 × 10⁻³ mol L⁻¹, the limiting molecular area of lecithin increased from 110.5 to 138.5 Å², but the collapse pressure of the monolayer decreased from 47.6 to 42.3 mN m⁻¹, indicating the destabilization of lipid monolayer caused by the penetration of tryptophan. The resistance spectra of s-BLM demonstrate that the existence of tryptophan leads to the formation of some defects in s-BLM and the destabilization of s-BLM. The values of electron-transfer resistance and double layer capacitance respectively decreased from 5.765 × 10⁶ Ω and 3.573 × 10⁻⁸ F to 1.391 × 10⁶ Ω and 3.340 × 10⁻⁸ F when the concentration of tryptophan increased from 0 to 2 × 10⁻³ mol L⁻¹. Correspondingly, the breakdown voltage of s-BLM decreased from 2.51 to 1.72 V.     

Electrocatalytic Oxidation of Tryptophan at Gold Nanoparticle‐Modified Carbon Ionic Liquid Electrode

The electrochemical behavior of tryptophan was studied at the carbon ionic liquid electrode (CILE) modified with gold nanoparticle (GNP). This electrode has a stable and excellent response toward tryptophan. Under optimum experimental conditions, the calibration curve was linear in the tryptophan concentration range of 5 to 900 µM with an excellent correlation coefficient (0.995). The experimental limit of detection was 4 µM. Contrary to many other electrodes, the oxidation of tryptophan on GNP/CILE does not result in electrode fouling. GNP/CILE has been effectively applied to the determination of tryptophan in composite amino acid injection.