22 research outputs found
Methods for Assessing Nuclear Rotation and Nuclear Positioning in Developing Skeletal Muscle Cells
Quantitative Assessment of Protein Structural Models by Comparison of H/D Exchange MS Data with Exchange Behavior Accurately Predicted by DXCOREX
Optogenetic approach for targeted activation of global calcium transients in differentiated C2C12 myotubes
The Public Health Perspective in Health Promotion and Disability Prevention for Older Adults: The Role of the Centers for Disease Control and Prevention
The Nucleotide Exchange Factor Ric-8A Is a Chaperone for the Conformationally Dynamic Nucleotide-Free State of Gαi1
Real-time HD Exchange Kinetics of Proteins from Buffered Aqueous Solution with Electrothermal Supercharging and Top-Down Tandem Mass Spectrometry
Electrothermal supercharging (ETS) with electrospray ionization produces highly charged protein ions from buffered aqueous solutions in which proteins have native folded structures. ETS increases the charge of ribonuclease A by 34% whereas only a 6% increase in charge occurs for a reduced alkylated form of this protein, which is unfolded and ~66% random coil in this solution. These results indicate that protein denaturation that occurs in the ESI droplets is the primary mechanism for ETS. ETS does not affect the extent of solution-phase hydrogen-deuterium exchange (HDX) that occurs for four proteins that have significantly different structures in solution, consistent with a droplet lifetime that is significantly shorter than observable rates of HDX. Rate constants for HDX of ubiquitin are obtained with a spatial resolution of ~1.3 residues with ETS and electron transfer dissociation of the 10+ charge-state using a single capillary containing a few ÎĽL of protein solution in which HDX continuously occurs. HDX protection at individual residues with ETS HDX is similar to that with reagent supercharging HDX and with solution phase NMR, indicating that the high spray potentials required to induce ETS do not lead to HD scrambling