muramidase and cadmium-telluride

The effect of 3-mercaptopropionic acid (MPA)-capped CdTe quantum dots (QDs) on lysozyme was systematically investigated by spectroscopic methods, enzyme activity assay, and calorimetry techniques. Results show that the MPA-capped CdTe QDs binded to lysozyme through van der Walls forces and hydrogen bondings, causing the decrement of α-helical content (∼7%) and increment of β-sheet content (∼11%) of lysozyme. The binding caused static quenching of the fluorescence, while the microenvironment of aromatic amino acid residues did not show any significant alteration. The lysozyme activity was affected by the increasing exposure of QDs, it was inhibited to 53.77% under a 6 × 10

Topics: 3-Mercaptopropionic Acid; Animals; Cadmium Compounds; Calorimetry, Differential Scanning; Chickens; Fluorescence; Hydrogen Bonding; Muramidase; Protein Structure, Secondary; Quantum Dots; Structure-Activity Relationship; Tellurium

The effect of N-acetyl-l-cysteine-capped CdTe quantum dots (NAC-CdTe QDs) with different sizes on lysozyme was investigated by isothermal titration calorimetry (ITC), enzyme activity assays, and multi-spectroscopic methods. ITC results proved that NAC-CdTe QDs can spontaneously bind with lysozyme and hydrophobic force plays a major role in stabilizing QDs-lysozyme complex. Multi-spectroscopic measurements revealed that NAC-CdTe QDs caused strong quenching of the lysozyme's fluorescence in a size-dependent quenching manner. Moreover, the changes of secondary structure and microenvironment in lysozyme caused by the NAC-CdTe QDs were higher with a bigger size. The results of enzyme activity assays showed that the interaction between lysozyme and NAC-CdTe QDs inhibited the activity of lysozyme and the inhibiting effect was in a size-dependent manner. Based on these results, we conclude that NAC-CdTe QDs with larger particle size had a larger impact on the structure and function of lysozyme.

Topics: Acetylcysteine; Cadmium Compounds; Catalytic Domain; Enzyme Inhibitors; Muramidase; Particle Size; Protein Binding; Protein Structure, Secondary; Quantum Dots; Tellurium; Thermodynamics

Lysozyme with a small monomeric globular enzymatic protein is part of the innate immune system, and its deficiency can cause the increased incidence of disease. Herein, we devise a new signal-enhanced fluorescence aptasensing platform for quantitative screening of lysozyme by coupling with rolling circle amplification (RCA) and strand hybridization reaction, accompanying the assembly of CdTe/CdSe quantum dots (QDs) and hemin/G-quadruplex DNzyme. Initially, target-triggered release of the primer was carried out from DNA duplex via the reaction of the aptamer with the analyte, and the released primer could be then utilized as the template to produce numerous repeated oligonucleotide sequences by the RCA reaction. Following that, the formed long-stranded DNA simultaneously hybridized with the CdTe/CdSe QD-labeled probe and hemin/G-quadruplex DNzyme strand in the system, thereby resulting in the quenching of QD fluorescent signal through the proximity hemin/G-quadruplex DNzyme on the basis of transferring photoexcited conduction band electrons of quantum dots to Fe(III)/Fe(II)-protoporphyrin IX (hemin) complex. Under optimal conditions, the fluorescent signal decreased with the increasing target lysozyme within the dynamic range from 5.0 to 500nM with a detection limit (LOD) of 2.6nM at the 3s

Topics: Aptamers, Nucleotide; Biosensing Techniques; Cadmium Compounds; DNA, Catalytic; Fluorescent Dyes; G-Quadruplexes; Hemin; Humans; Limit of Detection; Muramidase; Nucleic Acid Amplification Techniques; Quantum Dots; Selenium Compounds; Spectrometry, Fluorescence; Tellurium

In this work, we developed an ultrasensitive "turn on-off" fluorescence nanosensor for lysozyme (Lyz) detection. The novel nanosensor was constructed with the carboxymethyl chitosan modified CdTe quantum dots (CMCS-QDs). Firstly, the CMCS-QDs were fabricated via the electrostatic interaction between amino groups in CMCS polymeric chains and carboxyl groups on the surface of QDs. In the fluorescence "turn-on" step, the strong binding ability between Zn(2+) and CMCS on the surface of QDs can enhance the photoluminescence intensity (PL) of QDs. In the following fluorescence "turn-off" step, the N-acetyl-glucosamine (NAG) section along the CMCS chains was hydrolyzed by Lyz. As a result, Zn(2+) was released from the surface of QDs, and the Lyz-QDs complexes were formed to quench the QDs PL. Under the optimal conditions, there was a good linear relationship between the PL of QDs and the Lyz concentration (0.1-1.2 ng/mL) with the detection limit of 0.031 ng/mL. The developed method was ultrasensitive, highly selective and fast. It has been successfully employed in the detection of Lyz in the serum with satisfactory results.

Topics: Biosensing Techniques; Cadmium Compounds; Chitosan; Fluorescence; Humans; Limit of Detection; Muramidase; Quantum Dots; Spectrometry, Fluorescence; Tellurium; Zinc

Water-soluble cysteamine (CA) capped CdTe quantum dots (QDs) conjugated with lysozyme binding DNA (LBD) was constructed for luminescent sensing of lysozyme by forming a ternary self-assembly complex. Addition of negatively charged lysozyme binding DNA to the positively charged CA capped CdTe QDs buffer solution (Tris-HCl pH 7.4) could lead to the formation of QDs-LBD complex through electrostatic interactions. Once lysozyme was introduced into the CdTe QDs-LBD system, it could bind specifically with the QDs-LBD complex, resulting in fluorescence emission enhancement of the QDs due to the surface inert of QDs. At a given amount of LBD and CdTe QDs (LBD: QDs=2: 1), the fluorescence intensity enhancement of QDs was linear with lysozyme concentration over the range of 8.9-71.2 nM, with a detection limit of 4.3 nM. Due to the specific binding of LBD with lysozyme, this approach displayed high selectivity for lysozyme recognition. The sensing mechanism was confirmed by DLS and zeta potential measurement, and agarose gel electrophoresis experiment. Furthermore, the proposed CA-capped CdTe QDs-LBD sensor was applied to lysozyme detection in mouse serum and human morning urine samples, which showed high sensitivity and selectivity in the complex biological sample.

Topics: Amino Acids; Animals; Anisotropy; Biosensing Techniques; Cadmium Compounds; Cysteamine; DNA; Electrophoresis, Agar Gel; Fluorescent Dyes; Humans; Limit of Detection; Mice; Muramidase; Nanotechnology; Quantum Dots; Sensitivity and Specificity; Spectrometry, Fluorescence; Tellurium; Urinalysis

The interactions of lysozyme and calf thymus DNA (ctDNA) or thioglycolic acid (TGA) modified CdTe nanoparticles in aqueous solution have been studied by resonance light-scattering (RLS) spectroscopy. At pH 7.2 Britton-Robinson (BR) buffer solution and pH 7.4 phosphate buffered saline (PBS), the RLS signals of ctDNA and TGA modified CdTe nanoparticles were greatly enhanced by lysozyme in the region of 220-750 nm characterized by the peak around 306 and 353 nm, respectively. Under optimal conditions, the increase of RLS intensity of the two systems is proportional to the concentration of lysozyme. The linear range is 0.1-25 microg/ml for the lysozyme-ctDNA system, and 0.2-10.7 microg/ml for the lysozyme-TGA modified CdTe nanoparticles system. The detection limit is 0.041 microg/ml for the lysozyme-ctDNA system, and 0.083 microg/ml for the lysozyme-TGA modified CdTe nanoparticles system, respectively. Meanwhile lysozyme can also be used as a probe to determine the ctDNA. The increase of RLS intensity of the system is also proportional to the concentration of ctDNA. The linear range is 0.078-13 microg/ml. The detection limit is 0.024 microg/ml. Three kinds of samples were analyzed with satisfactory results.

Topics: Cadmium Compounds; DNA; Light; Microscopy, Electron, Transmission; Muramidase; Nanoparticles; Scattering, Radiation; Tellurium

Nanoparticles of cadmium telluride (CdTe) coated with thioglycolic acid (TGA) were prepared in the water phase. The interaction between CdTe nanoparticles (NPs) and lysozyme (Lyz) was investigated by fluorescence and circular dichroism (CD) spectroscopy at pH 7.40. It was proved that the fluorescence quenching of Lyz by CdTe NPs was mainly a result of the formation of CdTe-Lyz complex. By the fluorescence quenching results, the Stern-Volmer quenching constant (K(SV)), binding constant (Ka) and binding sites (n) were calculated. The binding distance (r) between Lyz (the donor) and CdTe NPs (the acceptor) was obtained according to fluorescence resonance energy transfer (FRET). Gradual addition of CdTe NPs to the solution of Lyz led to a marked increase in fluorescence polarization (P) of Lyz, which indicated that CdTe NPs were located in a restricted environment of Lyz. The effect of CdTe NPs on the conformation of Lyz has been analyzed by means of synchronous fluorescence spectra and CD spectra, which provided the evidence that the secondary structure of Lyz has been changed by the interaction of CdTe NPs with Lyz.

Topics: Binding Sites; Cadmium Compounds; Circular Dichroism; Glycolates; Kinetics; Models, Statistical; Muramidase; Nanoparticles; Nanotechnology; Protein Binding; Protein Conformation; Protein Structure, Secondary; Spectrometry, Fluorescence; Spectrophotometry; Tellurium; Water

Nanoparticles of cadmium telluride coated with mercaptoacetic acid were prepared in the water phase. Further, an assay of lysozyme with a sensitivity at the nanogram level is proposed. At pH 7.28, lysozyme with positive charges can interact with CdTe nanoparticles. The resonance light-scattering (RLS) signals of functionalized nano-CdTe were greatly enhanced by lysozyme in the region of 300-600 nm, characterized with peaks located at 367, 470 and 533 nm. A linear relationship could be established between the enhanced RLS intensity and the lysozyme concentration in the range of 0.06-4.0 microg mL-1. The limit of detection was 9.5 ng mL-1. The contents of lysozyme were determined with recoveries of 95.6-104.8% and RSD of 1.5-2.3%, respectively. This method is sensitive, rapid, accurate and simple, and provides a new and reliable means for the quantity determination of lysozyme.

Topics: Buffers; Cadmium Compounds; Colloids; Hydrogen-Ion Concentration; Muramidase; Nanoparticles; Osmolar Concentration; Tellurium; Time Factors