Gas-phase Förster resonance energy transfer (FRET) combines the advantages of mass spectrometry a... more Gas-phase Förster resonance energy transfer (FRET) combines the advantages of mass spectrometry and fluorescence spectroscopy for the conformational analysis of mass-selected biomolecules. While this implementation of FRET in the gas phase promises detailed insights for fundamental and applied studies, the gas-phase environment also poses great challenges. For FRET, fluorophore pairs are typically covalently attached to strategic binding sites in the backbone of a biomolecule, using short linkers. The linker further increases the mobility of the dye, contributing to rotational averaging of the relative orientation of the transition dipole moments of donor and acceptor. However, little is known about the fluorophore’s degrees of freedom in the gas phase and how it may be influenced by intramolecular interactions. In this study, we test the influence of a fluorophore’s linker length on the measured FRET efficiencies in the gas phase to probe the mobility of the fluorophore. An increas...
Laser-induced fluorescence studies on mass-selected biomolecules are a promising route to underst... more Laser-induced fluorescence studies on mass-selected biomolecules are a promising route to understand their properties in the gas phase and probe their intrinsic properties in a solvent-free environment. Fluorescence has been used to investigate the conformation and dynamics of gaseous biomolecular ions. With Förster Resonance Energy Transfer (FRET), it is now possible to obtain sensitive intramolecular distance information from large biomolecules, like proteins, with high chemical specificity. With growing interest and applications, gas-phase fluorescence measurements can shed greater light on the characteristics of proteins in the gas phase. Compared to the solution phase measurements, gas-phase fluorescence can also help understand the influence of solvent interactions on the protein structure and function.
Internal combustion engine occupies a significant spot in present day. with the most recent plan ... more Internal combustion engine occupies a significant spot in present day. with the most recent plan alteration in different part being advanced to improve the efficiency and generally speaking performance, one segment has been kept untouched that is camshaft. Cam control the breathing channels of the engine that is the planning of valves through which fuel air mixture enters and fumes is driven out. with help of push rods , rocker arms, stiff springs. With increasing performance demands engine specialists and researchers over the world are perusing radical cam less structure which promises to give ICEs a greater improvement in effectiveness. The fantasy about accomplishing cam less technology is shutting reality at rapid rate. With cam less technology put into practice, the engine power can be improved, pollutant emissions could be controlled and better mileage is acquired, in addition to that the whole mechanism of pulley or belt drive is replaced by the camless engine with greater ef...
Additional file 5: A. SDS-PAGE analysis of enzyme immobilization. B. Table showing enzyme activit... more Additional file 5: A. SDS-PAGE analysis of enzyme immobilization. B. Table showing enzyme activity in immobilization.
Additional file 3: 3.1. Gene designing strategy of rc-TGL. 3.2. PCR and cloning of rc-TGL into pE... more Additional file 3: 3.1. Gene designing strategy of rc-TGL. 3.2. PCR and cloning of rc-TGL into pET23a expression vector. 3.3. Primer sequences used to construct rc-TGL.
Path finder was send to Mars in 1998.The metal detector concerns with the radio frequency. This r... more Path finder was send to Mars in 1998.The metal detector concerns with the radio frequency. This robot is just like a prototype for the Path finder. The project is controlled by the remote and robot moved in forward, backwards, left, right direction by geared motors. A camera is on the receiver side to see the images surrounding the objects. When the receiver is moving on the land the circuit produces the beep sound when metal is detected. The sound produced will be transmitted to the transmitter part. User can monitor the alarms on television and view the images. INTRODUCTION: The object of the transmitter ckt is to control the position and moving of robot to detect metals and see the images around the robot. The aim of the project is to use the radio frequency bands. The moving of robot is regulated by radio signals through air. The receiver receives the signal and movement of robot is regulated. It consists of transmitter and receiver part. Transmitter part consists of encoder swi...
Attendance marking in a classroom is a very time consuming task. It is a very difficult for lectu... more Attendance marking in a classroom is a very time consuming task. It is a very difficult for lecturers to take attendance in a class of very large number of students. This also reduces the time of lecture. This paper presents Automatic Attendance System using Face detection of students in class using MATLAB . This syste m uses Viola-Jones algorithm to detect the faces from the images captured by camera. These images are compared using SURF matching algorithm with the stored images of students. These two algorithms are implemented in MATLAB. The system can be operated automatically or manually. The focus is to make a fully automatic system which works on basis of time-table of class-room. We make a standalone application for this automatic attendance system which can work on any 64-bit computer with no need of MATLAB software.
Gas-phase fluorescence spectroscopy is still in its infancy, which demands further instrumental d... more Gas-phase fluorescence spectroscopy is still in its infancy, which demands further instrumental developments. In this work, a Fourier-transform ion cyclotron resonance mass spectrometer (FT-ICR MS), equipped with a lab-developed data acquisition (DAQ) system, was coupled to a tunable femtosecond (fs) laser and a state-of-the-art optical system for fluorescence studies of mass-selected ions. For excitation, a laser beam was focused (beam size <1.0 mm) into the cylindrical ICR cell. A wire mesh replaced the back trapping plate, allowing ~10% of the fluorescence emitted from trapped ions to be collected by a lens installed beside the wire mesh. The collected fluorescence light was then transmitted outside of the mass spectrometer via fiber optics. A novel accumulation during detection (ADD) scheme was developed to increase the duty cycle of gas-phase fluorescence spectroscopy experiments. With ADD, >90% duty cycle for mass spectrometric and fluorescence experiments could be achieved. This instrument was able to perform fluorescence experiments on various ions, from simple rhodamine dyes to large biomolecules (i.e., peptide and protein) labeled with dyes of various optical properties. A fluorescence lifetime measurement of trapped rhodamine 6G cations was also acquired, yielding a value of 5.97 ± 0.23 ns. This setup has a broad mass range and decent fluorescence spectroscopy performance (i.e., a fluorescence spectrum of rhodamine 6G can be acquired with good S/N in a minute). Finally, this setup also allows more challenging gas-phase fluorescence spectroscopy experiments, e.g., of low quantum yield fluorophores and large biomolecules in their native state that appear at high m/z, which may not be doable with quadrupole ion traps (QITs).
exclusion on the accessible volumes. To investigate whether this selfassociative behavior applies... more exclusion on the accessible volumes. To investigate whether this selfassociative behavior applies across a broader range of species and other model organisms, we examined Xenopus laevis Arr1 (xArr1) which bears 68% sequence identity to mammalian Arr1s. Homology modeling based on the bovine crystal structure suggests that it is likely to have a very similar tertiary structure. Residues of mammalian Arr1 hypothesized to be at dimer and tetramer interfaces by their ability to affect the extent of oligomerization are mostly conserved in X. laevis as well, suggesting xArr1 would likely follow the same self-association pattern. We performed analytical ultracentrifugation experiments on xArr1 as well as bovine Arr1 (bArr1) for comparison. Our data for bArr1 agrees well with K D s from published studies, and the K D dimer for xArr1 fits well into the range published for human, mouse, and bovine Arr1. Surprisingly, at concentrations up to 210 micromolar, our results show xArr1 as monomer and dimer only, with no evidence of higher order oligomers. The mechanism underlying this difference is under investigation.
This page was generated automatically upon download from the ETH Zurich Research Collection. For ... more This page was generated automatically upon download from the ETH Zurich Research Collection. For more information, please consult the Terms of use. Fluorescence-based detection of desolvated protein ions generated in an aqueous electrospray plume at ambient conditions
Advances in Experimental Medicine and Biology, 2018
This article begins with a general review of cell adhesion molecules (CAMs) and narrows the focus... more This article begins with a general review of cell adhesion molecules (CAMs) and narrows the focus down progressively to the cadherins (calcium binding-dependent CAMs), classifications of subfamilies of the cadherins, type I (E- and N-) cadherins, evolutionary relationships amongst cadherins, structural-mechanical and functional consequences of calcium binding to the cadherins, differential molecular interactions involving the extracellular (ecto) and intracellular (cytoplasmic) domains of the cadherins, multiple adherence-related homophilic and heterophilic interactions and associated functions of E- and N-cadherin in organismal development and disease and cadherin trafficking and membrane rafts. It ends by summarizing multiple perspectives and hypotheses concerning different aspects of cadherin structure, stability and function.
Myosin II is the motor protein responsible for contractility in muscle and nonmuscle cells 1. The... more Myosin II is the motor protein responsible for contractility in muscle and nonmuscle cells 1. The molecule has two identical heads attached to an elongated tail, and can exist in two conformations: 10S and 6S, named for their sedimentation coefficients 2,3. The 6S conformation has an extended tail and assembles into polymeric filaments, which pull on actin filaments to generate force and motion. In 10S myosin, the tail is folded into three segments and the heads bend back and interact with each other and the tail 3-7 , creating a compact conformation, in which ATPase activity, actin activation and filament assembly are all highly inhibited 7,8. This switched-off structure appears to function as a key energy-conserving storage molecule in muscle and nonmuscle cells 9-12 , which can be activated to form functional filaments as needed 13-but the mechanism of its inhibition is not understood. Here we have solved the structure of smooth muscle 10S myosin to a global resolution of 4.3 Å by cryo-EM, revealing near-atomic level details of its structure for the first time. The reconstruction provides a new understanding of the head and tail regions of the molecule and of the key intramolecular contacts that cause inhibition. Our results suggest an atomic model for the off-state of myosin II, for its activation and unfolding by phosphorylation, and for understanding the clustering of disease-causing mutations near sites of intramolecular interaction. Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:
Gas-phase Förster resonance energy transfer (FRET) combines the advantages of mass spectrometry a... more Gas-phase Förster resonance energy transfer (FRET) combines the advantages of mass spectrometry and fluorescence spectroscopy for the conformational analysis of mass-selected biomolecules. While this implementation of FRET in the gas phase promises detailed insights for fundamental and applied studies, the gas-phase environment also poses great challenges. For FRET, fluorophore pairs are typically covalently attached to strategic binding sites in the backbone of a biomolecule, using short linkers. The linker further increases the mobility of the dye, contributing to rotational averaging of the relative orientation of the transition dipole moments of donor and acceptor. However, little is known about the fluorophore’s degrees of freedom in the gas phase and how it may be influenced by intramolecular interactions. In this study, we test the influence of a fluorophore’s linker length on the measured FRET efficiencies in the gas phase to probe the mobility of the fluorophore. An increas...
Laser-induced fluorescence studies on mass-selected biomolecules are a promising route to underst... more Laser-induced fluorescence studies on mass-selected biomolecules are a promising route to understand their properties in the gas phase and probe their intrinsic properties in a solvent-free environment. Fluorescence has been used to investigate the conformation and dynamics of gaseous biomolecular ions. With Förster Resonance Energy Transfer (FRET), it is now possible to obtain sensitive intramolecular distance information from large biomolecules, like proteins, with high chemical specificity. With growing interest and applications, gas-phase fluorescence measurements can shed greater light on the characteristics of proteins in the gas phase. Compared to the solution phase measurements, gas-phase fluorescence can also help understand the influence of solvent interactions on the protein structure and function.
Internal combustion engine occupies a significant spot in present day. with the most recent plan ... more Internal combustion engine occupies a significant spot in present day. with the most recent plan alteration in different part being advanced to improve the efficiency and generally speaking performance, one segment has been kept untouched that is camshaft. Cam control the breathing channels of the engine that is the planning of valves through which fuel air mixture enters and fumes is driven out. with help of push rods , rocker arms, stiff springs. With increasing performance demands engine specialists and researchers over the world are perusing radical cam less structure which promises to give ICEs a greater improvement in effectiveness. The fantasy about accomplishing cam less technology is shutting reality at rapid rate. With cam less technology put into practice, the engine power can be improved, pollutant emissions could be controlled and better mileage is acquired, in addition to that the whole mechanism of pulley or belt drive is replaced by the camless engine with greater ef...
Additional file 5: A. SDS-PAGE analysis of enzyme immobilization. B. Table showing enzyme activit... more Additional file 5: A. SDS-PAGE analysis of enzyme immobilization. B. Table showing enzyme activity in immobilization.
Additional file 3: 3.1. Gene designing strategy of rc-TGL. 3.2. PCR and cloning of rc-TGL into pE... more Additional file 3: 3.1. Gene designing strategy of rc-TGL. 3.2. PCR and cloning of rc-TGL into pET23a expression vector. 3.3. Primer sequences used to construct rc-TGL.
Path finder was send to Mars in 1998.The metal detector concerns with the radio frequency. This r... more Path finder was send to Mars in 1998.The metal detector concerns with the radio frequency. This robot is just like a prototype for the Path finder. The project is controlled by the remote and robot moved in forward, backwards, left, right direction by geared motors. A camera is on the receiver side to see the images surrounding the objects. When the receiver is moving on the land the circuit produces the beep sound when metal is detected. The sound produced will be transmitted to the transmitter part. User can monitor the alarms on television and view the images. INTRODUCTION: The object of the transmitter ckt is to control the position and moving of robot to detect metals and see the images around the robot. The aim of the project is to use the radio frequency bands. The moving of robot is regulated by radio signals through air. The receiver receives the signal and movement of robot is regulated. It consists of transmitter and receiver part. Transmitter part consists of encoder swi...
Attendance marking in a classroom is a very time consuming task. It is a very difficult for lectu... more Attendance marking in a classroom is a very time consuming task. It is a very difficult for lecturers to take attendance in a class of very large number of students. This also reduces the time of lecture. This paper presents Automatic Attendance System using Face detection of students in class using MATLAB . This syste m uses Viola-Jones algorithm to detect the faces from the images captured by camera. These images are compared using SURF matching algorithm with the stored images of students. These two algorithms are implemented in MATLAB. The system can be operated automatically or manually. The focus is to make a fully automatic system which works on basis of time-table of class-room. We make a standalone application for this automatic attendance system which can work on any 64-bit computer with no need of MATLAB software.
Gas-phase fluorescence spectroscopy is still in its infancy, which demands further instrumental d... more Gas-phase fluorescence spectroscopy is still in its infancy, which demands further instrumental developments. In this work, a Fourier-transform ion cyclotron resonance mass spectrometer (FT-ICR MS), equipped with a lab-developed data acquisition (DAQ) system, was coupled to a tunable femtosecond (fs) laser and a state-of-the-art optical system for fluorescence studies of mass-selected ions. For excitation, a laser beam was focused (beam size <1.0 mm) into the cylindrical ICR cell. A wire mesh replaced the back trapping plate, allowing ~10% of the fluorescence emitted from trapped ions to be collected by a lens installed beside the wire mesh. The collected fluorescence light was then transmitted outside of the mass spectrometer via fiber optics. A novel accumulation during detection (ADD) scheme was developed to increase the duty cycle of gas-phase fluorescence spectroscopy experiments. With ADD, >90% duty cycle for mass spectrometric and fluorescence experiments could be achieved. This instrument was able to perform fluorescence experiments on various ions, from simple rhodamine dyes to large biomolecules (i.e., peptide and protein) labeled with dyes of various optical properties. A fluorescence lifetime measurement of trapped rhodamine 6G cations was also acquired, yielding a value of 5.97 ± 0.23 ns. This setup has a broad mass range and decent fluorescence spectroscopy performance (i.e., a fluorescence spectrum of rhodamine 6G can be acquired with good S/N in a minute). Finally, this setup also allows more challenging gas-phase fluorescence spectroscopy experiments, e.g., of low quantum yield fluorophores and large biomolecules in their native state that appear at high m/z, which may not be doable with quadrupole ion traps (QITs).
exclusion on the accessible volumes. To investigate whether this selfassociative behavior applies... more exclusion on the accessible volumes. To investigate whether this selfassociative behavior applies across a broader range of species and other model organisms, we examined Xenopus laevis Arr1 (xArr1) which bears 68% sequence identity to mammalian Arr1s. Homology modeling based on the bovine crystal structure suggests that it is likely to have a very similar tertiary structure. Residues of mammalian Arr1 hypothesized to be at dimer and tetramer interfaces by their ability to affect the extent of oligomerization are mostly conserved in X. laevis as well, suggesting xArr1 would likely follow the same self-association pattern. We performed analytical ultracentrifugation experiments on xArr1 as well as bovine Arr1 (bArr1) for comparison. Our data for bArr1 agrees well with K D s from published studies, and the K D dimer for xArr1 fits well into the range published for human, mouse, and bovine Arr1. Surprisingly, at concentrations up to 210 micromolar, our results show xArr1 as monomer and dimer only, with no evidence of higher order oligomers. The mechanism underlying this difference is under investigation.
This page was generated automatically upon download from the ETH Zurich Research Collection. For ... more This page was generated automatically upon download from the ETH Zurich Research Collection. For more information, please consult the Terms of use. Fluorescence-based detection of desolvated protein ions generated in an aqueous electrospray plume at ambient conditions
Advances in Experimental Medicine and Biology, 2018
This article begins with a general review of cell adhesion molecules (CAMs) and narrows the focus... more This article begins with a general review of cell adhesion molecules (CAMs) and narrows the focus down progressively to the cadherins (calcium binding-dependent CAMs), classifications of subfamilies of the cadherins, type I (E- and N-) cadherins, evolutionary relationships amongst cadherins, structural-mechanical and functional consequences of calcium binding to the cadherins, differential molecular interactions involving the extracellular (ecto) and intracellular (cytoplasmic) domains of the cadherins, multiple adherence-related homophilic and heterophilic interactions and associated functions of E- and N-cadherin in organismal development and disease and cadherin trafficking and membrane rafts. It ends by summarizing multiple perspectives and hypotheses concerning different aspects of cadherin structure, stability and function.
Myosin II is the motor protein responsible for contractility in muscle and nonmuscle cells 1. The... more Myosin II is the motor protein responsible for contractility in muscle and nonmuscle cells 1. The molecule has two identical heads attached to an elongated tail, and can exist in two conformations: 10S and 6S, named for their sedimentation coefficients 2,3. The 6S conformation has an extended tail and assembles into polymeric filaments, which pull on actin filaments to generate force and motion. In 10S myosin, the tail is folded into three segments and the heads bend back and interact with each other and the tail 3-7 , creating a compact conformation, in which ATPase activity, actin activation and filament assembly are all highly inhibited 7,8. This switched-off structure appears to function as a key energy-conserving storage molecule in muscle and nonmuscle cells 9-12 , which can be activated to form functional filaments as needed 13-but the mechanism of its inhibition is not understood. Here we have solved the structure of smooth muscle 10S myosin to a global resolution of 4.3 Å by cryo-EM, revealing near-atomic level details of its structure for the first time. The reconstruction provides a new understanding of the head and tail regions of the molecule and of the key intramolecular contacts that cause inhibition. Our results suggest an atomic model for the off-state of myosin II, for its activation and unfolding by phosphorylation, and for understanding the clustering of disease-causing mutations near sites of intramolecular interaction. Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:
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