Ca(2+) -complex stability of GAPAGPLIVPY peptide in gas and aqueous phase, investigated by affinity capillary electrophoresis and molecular dynamics simulations and compared to mass spectrometric results.

Strong, sequence-specific gas-phase bindings between proline-rich peptides and alkaline earth metal ions in nanoESI-MS experiments were reported by Lehmann et al. (Rapid Commun. Mass Spectrom.

Optimization of affinity capillary electrophoresis for routine investigations of protein-metal ion interactions.

To facilitate the implementation of affinity capillary electrophoresis into routine binding screening studies of proteins with metal ions, method acceleration, transfer and precision improvement were investigated. Affinity capillary electrophoresis was accelerated by using shorter capillaries, employing lower sample concentrations and smaller injection volumes. Intra- and inter-instrument method transfers were investigated considering the temperature setting of the capillary cooling system.

A comprehensive platform to investigate protein-metal ion interactions by affinity capillary electrophoresis.

In this work, the behavior of several metal ions with different globular proteins was investigated by affinity capillary electrophoresis. Screening was conducted by applying a proper rinsing protocol developed by our group. The use of 0.

Precise, fast, and flexible determination of protein interactions by affinity capillary electrophoresis: part 3: anions.

The binding of physiologically anionic species or negatively charged drug molecules to proteins is of great importance in biochemistry and medicine. Since affinity capillary electrophoresis (ACE) has already proven to be a suitable analytical tool to study the influence of ions on proteins, this technique was applied here for comprehensively studying the influence of various anions on proteins of BSA, β-lactoglobulin, ovalbumin, myoglobin, and lysozyme. The analysis was performed using different selected anions of succinate, glutamate, phosphate, acetate, nitrate, iodide, thiocyanate, and pharmaceuticals (salicylic acid, aspirin, and ibuprofen) that exist in the anionic form at physiological pH 7.

Precise, fast and flexible determination of protein interactions by affinity capillary electrophoresis. Part 2: cations.

The influence of various cations as metal ions (barium, calcium, copper, magnesia, manganese, and nickel), pharmaceuticals (ephedrine, ethambutol, pilocarpine, and pirenzepine), arginine, and guanidine has been tested on BSA, β-lactoglobulin, and ovalbumin. Influences on proteins regarding changes in size, charge, or mass were of particular interest. ACE proved to be a suitable method to investigate these effects.

Precise, fast, and flexible determination of protein interactions by affinity capillary electrophoresis: part 1: performance.

A generic screening approach was performed to investigate the binding of various potential ligands to proteins in order to investigate how proteins interact with ions and the complete surrounding solution. This also allows to study binding behavior and its regulation and protein functionality in general in a native environment. The ACE technique affords an excellent precision by applying appropriate rinsing procedures using a pressure of 2.

The challenge to quantify proteins with charge trains due to isoforms or conformers.

Single proteins separated by 2-DE often show multiple spots spreading along the first dimension. In many cases, such charge trains are explained by isoform differences or by putative post-translational modifications including phosphorylation, glycosylation and others. We now report that individual spots of such charge trains on 2-D gels in fact often represent the same protein, but, apparently due to conformational changes, segregate to different isoelectric points.

Quantitative gel electrophoresis: new records in precision by elaborated staining and detection protocols.

Gel electrophoresis (GE) is a very common analytical technique for proteome research and protein analysis. Despite being developed decades ago, there is still a considerable need to improve its precision. Using the fluorescence of Colloidal Coomassie Blue -stained proteins in near-infrared (NIR), the major error source caused by the unpredictable background staining is strongly reduced.