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Article
Protein-Ligand Interaction by Ligand Titration, Fast Photochemical Oxidation of Proteins and Mass Spectrometry: LITPOMS
We report a novel method named LITPOMS (ligand titration, fast photochemical oxidation of proteins and mass spectrometry) to characterize protein-ligand binding stoichiometry, binding sites, and site-specific ...
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Article
Incorporation of a Reporter Peptide in FPOP Compensates for Adventitious Scavengers and Permits Time-Dependent Measurements
Incorporation of a reporter peptide in solutions submitted to fast photochemical oxidation of proteins (FPOP) allows for the correction of adventitious scavengers and enables the normalization and comparison o...
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Article
Protein Footprinting by Carbenes on a Fast Photochemical Oxidation of Proteins (FPOP) Platform
Protein footprinting combined with mass spectrometry provides a method to study protein structures and interactions. To improve further current protein footprinting methods, we adapted the fast photochemical o...
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Article
Dosimetry Determines the Initial OH Radical Concentration in Fast Photochemical Oxidation of Proteins (FPOP)
Fast photochemical oxidation of proteins (FPOP) employs laser photolysis of hydrogen peroxide to give OH radicals that label amino acid side-chains of proteins on the microsecond time scale. A method for quant...
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Article
An Improved Fast Photochemical Oxidation of Proteins (FPOP) Platform for Protein Therapeutics
Unlike small-molecule drugs, the size and dynamics of protein therapeutics challenge existing methods for assessing their high order structures (HOS). To extend fast photochemical oxidation of proteins (FPOP) ...
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Article
Fast Photochemical Oxidation of Proteins (FPOP) Maps the Epitope of EGFR Binding to Adnectin
Epitope map** is an important tool for the development of monoclonal antibodies, mAbs, as therapeutic drugs. Recently, a class of therapeutic mAb alternatives, adnectins, has been developed as targeted biolo...
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Article
Mass Spectrometry Combinations for Structural Characterization of Sulfated-Steroid Metabolites
Steroid conjugates, which often occur as metabolites, are challenging to characterize. One application is female-mouse urine, where steroid conjugates serve as important ligands for the pheromone-sensing neuro...
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Article
Protein-peptide affinity determination using an H/D exchange dilution strategy: Application to antigen-antibody interactions
A new methodology using hydrogen/deuterium amide exchange (HDX) to determine the binding affinity of protein-peptide interactions is reported. The method, based on our previously established approach, protein ...
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Article
A tuning method for electrically compensated ion cyclotron resonance mass spectrometer traps
We describe a method for tuning electrically compensated ion cyclotron resonance (ICR) traps by tracking the observed cyclotron frequency of an ion cloud at different oscillation mode amplitudes. Although we h...
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Article
The gramicidin dimer shows both EX1 and EX2 mechanisms of H/D exchange
We describe the use of H/D amide exchange and electrospray ionization mass spectrometry to study, in organic solvents, the pentadecapeptide gramicidin as a model for protein self association. In methanol-OD, a...
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Article
An electrically compensated trap designed to eighth order for FT-ICR mass spectrometry
We present the design, guided by theory to eighth order, and the first evaluation of a Fourier transform ion cyclotron resonance (FT-ICR) compensated trap. The purpose of the new trap is to reduce effects of t...
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Article
Application of SIMSTEX to oligomerization of insulin analogs and mutants
The propensity of various insulins and their analogs to oligomerize was investigated by mass spectrometric methods including measurement of the relative abundances of oligomers in the gas phase and the kinetic...
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Article
Determination of affinity constants and response factors of the noncovalent dimer of gramicidin by electrospray ionization mass spectrometry and mathematical modeling
The dimerization of gramicidin, a 15-residue membrane peptide, in solution can be viewed as a model for protein-protein interactions. We reported previously that the dimer can be observed when electrosprayed f...
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Article
Evaluation of different combinations of gated trap**, RF-only mode and trap compensation for in-field MALDI Fourier transform mass spectrometry
MALDI, while providing advantages such as the ability to do in-depth and repeated exploration of the sample, challenges the existing performance capabilities of Fourier transform mass spectrometry (FTMS). The ...
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Article
Modeling data from titration, amide H/D exchange, and mass spectrometry to obtain protein-ligand binding constants
We recently reported a new method for quantification of protein-ligand interaction by mass spectrometry, titration and H/D exchange (PLIMSTEX) for determining the binding stoichiometry and affinity of a wide r...