Synthesis and expression of w-conotoxin MVIIA

Cai, Minying; Qu, Xun

doi:10.1007/0-306-46880-8_33

Cited by 5 publications

(9 citation statements)

References 7 publications

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“…Mobility-selected mass spectra were extracted with the instrument control software MassLynx V4. Extracted mass spectra were converted into .mgf (Mascot Generic Format) files and imported into Mash Explorer, 39 where spectra were deconvoluted by the eThrash algorithm 40 with a S/N threshold of 3, peak background ratio of 1, peptide minimum background ratio of 1, and minimum isotopic fit % of 80. The covalently modified and unmodified CID fragments obtained for all experiments were investigated against the ubiquitin primary sequence by applying custom PTMs equal to the mass of the covalent modification formed by the ion/ion reactions (i.e., 182.98 Da) at the N and C termini.…”

Section: Discussionmentioning

confidence: 99%

Ion Mobility and Gas-Phase Covalent Labeling Study of the Structure and Reactivity of Gaseous Ubiquitin Ions Electrosprayed from Aqueous and Denaturing Solutions

Carvalho

Kit

Webb

2020

J. Am. Soc. Mass Spectrom.

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Gas-phase ion/ion chemistry was coupled to ion mobility/mass spectrometry analysis to correlate the structure of gaseous ubiquitin to its solution structures with selective covalent structural probes. Collision cross section (CCS) distributions were measured to ensure the ubiquitin ions were not unfolded when they were introduced to the gas phase. Aqueous solutions stabilizing the native state of ubiquitin yielded folded ubiquitin structures with CCS values consistent with previously published literature. Denaturing solutions favored several families of unfolded conformations for most of the charge states evaluated. Gas-phase covalent labeling via ion/ion reactions was followed by collision induced dissociation of the intact, labeled protein to determine which residues were labeled. Ubiquitin 5 + and 6 + electrosprayed from aqueous conditions were covalently modified preferentially at the lysine 29 and arginine 54 positions, indicating that elements of three-dimensional structure were maintained in the gas phase. On the other hand, most ubiquitin ions produced in denaturing conditions were labeled at various other lysine residues, likely due to the availability of additional sites following methanol and low pHinduced unfolding. These data support the conservation of ubiquitin structural elements in the gas phase. The research presented here provides the basis for residue-specific characterization of biomolecules in the gas phase.

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Section: Discussionmentioning

confidence: 99%

Ion Mobility and Gas-Phase Covalent Labeling Study of the Structure and Reactivity of Gaseous Ubiquitin Ions Electrosprayed from Aqueous and Denaturing Solutions

Carvalho

Kit

Webb

2020

J. Am. Soc. Mass Spectrom.

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“…This section provides a brief summary of the algorithm for the 3D tomography reconstruction; more detailed discussions can be found in [27,28,35]. Tomography reconstruction can be summarized into two steps.…”

Section: D Tomography Reconstruction and Validationmentioning

confidence: 99%

“…The algorithm used here to solve for the 3D distribution was a modified variation of the Algebraic Reconstruction Technique (ART) [36]. The major modifications included the use of simulated annealing to improve the reconstruction [37,38] and the use of geometrical ray-tracing [27,39] and Monte Carlo simulation [40,41], so that the inversion can be performed on projections gathered from an arbitrary experimental setup (i.e., not restricted to the so-called parallel-beam projections). The tomographic algorithm used here has been validated via numerical simulations [27] and controlled experiments in both non-reactive [31] and reactive flows [26,27].…”

Section: D Tomography Reconstruction and Validationmentioning

confidence: 99%

“…The side view shown in Figure 4f is unable to reveal the 3D structure of the flame. Besides such visual comparison, the 3D tomographic reconstruction has also been validated via other methods in our past work, for both numerical simulations [27] to controlled flames [42].…”

Section: D Tomography Reconstruction and Validationmentioning

confidence: 99%

“…More specifically, with the TC diagnostic we have measured line-of-sight cavity chemiluminescence images (termed projections) simultaneously from 8 different orientations, each at a rate of 20 kHz. These projections were then fed into a tomographic reconstruction algorithm [27] as inputs to obtain 3D measurements of combustion processes. Processing the tomographic reconstruction frame by frame resulted in a sequence of 3D measurements at a rate of 20 kHz.…”

Section: Introductionmentioning

confidence: 99%

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From ignition to stable combustion in a cavity flameholder studied via 3D tomographic chemiluminescence at 20 kHz

Lei

et al. 2016

Combustion and Flame

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This work reports the study of the ignition processes in a Mach-2 cavity combustor based on three-dimensional (3D) measurements at 20 kHz. The 3D measurements were obtained by a combination of tomographic chemiluminescence and fiber-based endoscopes. Measurements of 3D flame and flow properties were reported under two fueling conditions of the combustor. The properties included 3D volume, surface area, shape factor, and 3D3C (three-dimensional and three-component) velocity of the ignition kernel. These results clearly distinguished the ignition stage from the stable combustion stage of the combustor and enabled the determination of a transition time to quantify both stages. The analysis of the change of the ignition kernel's shape, when combined with the 3D3C velocity measurements, also illustrated flame-flow interactions in the cavity combustor. These results demonstrated the utility of the 3D diagnostics to overcome some of the limitations of established planar diagnostics and to resolve the dynamics of highspeed combustion devices both spatially and temporally.

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A Top-Down Proteomics Platform Coupling Serial Size Exclusion Chromatography and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

et al. 2019

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Mass spectrometry (MS)-based top-down proteomics provides rich information about proteoforms arising from combinatorial amino acid sequence variations and post-translational modifications (PTMs). Fourier transform ion cyclotron resonance (FT-ICR) MS affords ultra-high resolving power and provides high-accuracy mass measurements, presenting a powerful tool for top-down MS characterization of proteoforms. However, detection and characterization of large proteins from complex mixtures remain challenging due to the exponential decrease in S:N with increasing molecular weight (MW) and co-eluting low-MW proteins; thus, size-based fractionation of complex protein mixtures prior to MS analysis is necessary. Here, we directly combine MScompatible serial size exclusion chromatography (sSEC) fractionation with 12 T FT-ICR MS for targeted top-down characterization of proteins from complex mixtures extracted from the human and swine heart proteome. Benefiting from the ultra-high resolving power of FT-ICR, we isotopically resolved 31 distinct proteoforms (30-50 kDa) simultaneously in a single mass spectrum within a 100 m/z window. Notably, within a 5 m/z window, we obtained baseline isotopic resolution for 6 distinct large proteoforms (30-50 kDa). The ultra-high resolving power of FT-ICR MS combined with sSEC fractionation enabled targeted top-down analysis of large proteoforms (>30 kDa) from the human heart proteome without extensive chromatographic separation or protein purification. Further separation of proteoforms inside of the mass spectrometer (in-MS) allowed for isolation of individual proteoforms for targeted electron capture dissociation (ECD) for high sequence coverage. sSEC/FT-ICR ECD facilitated identification and sequence characterization of important metabolic enzymes. This platform, which facilitates deep *

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Synthesis and expression of w-conotoxin MVIIA

Cited by 5 publications

References 7 publications

Ion Mobility and Gas-Phase Covalent Labeling Study of the Structure and Reactivity of Gaseous Ubiquitin Ions Electrosprayed from Aqueous and Denaturing Solutions

Ion Mobility and Gas-Phase Covalent Labeling Study of the Structure and Reactivity of Gaseous Ubiquitin Ions Electrosprayed from Aqueous and Denaturing Solutions

From ignition to stable combustion in a cavity flameholder studied via 3D tomographic chemiluminescence at 20 kHz

A Top-Down Proteomics Platform Coupling Serial Size Exclusion Chromatography and Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

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