Protein Folding Research
Background & Overview
Proteins are dynamic molecules that exhibit a wide range of behaviors and perform many functions in cells; understanding how they fold into complex three-dimensional (3D) structures and how these structures remain stable is critical to explaining their overall behavior. However, studying protein folding and stability is a difficult task. The complexity of the folding process and the diversity and interchangeability between possible protein conformations requires highly time-sensitive and complex experiments. BOC Sciences can use hydrogen/deuterium exchange (HDX) technology based on solvent exchange and protein engineering to study protein folding mechanisms and provide valuable information. You only need to send us your samples, and we will take care of all the follow-up matters of the project, including sample labeling, mass spectrometry detection, raw data analysis, etc.
Analysis Process
Fig.1 Typical workflow for continuous labeling coupled to “bottom-up” Hydrogen Deuterium eXchange Mass Spectrometry (HDX-MS).[1]
Our Services
BOC Sciences uses hydrogen-deuterium exchange mass spectrometry to study proteins in their folded or partially folded states and reports on the stability of the hydrogen bonds that exist in these states. Changing environmental conditions, such as pH or denaturant concentration, can also provide quantitative information on the level of protection of individual amide protons against protein conformational changes.
Our services include the determination of protein folding sequences, studies of single nucleotide polymorphisms and their impact on disease states, and exploration of mechanisms of protein aggregation and amyloid fibril formation, among others.
Through the use of molecular chaperones, cells are preprogrammed to deal with many types of protein folding and misfolding problems. Our hydrogen-deuterium exchange mass spectrometry technique is also very useful for studying how protein folding is altered by chaperones and how molecular chaperones function.
Fig.2 HDX MS in the study of chaperone-assisted protein folding.[2]
To better understand the changes in protein structure, we also used hydrogen-deuterium exchange mass spectrometry in combination with native mass spectrometry and chemical cross-linking.
Technical Advantages
The main advantages of HDX MS compared to NMR are the following:
- Relatively easy to study high molecular weight proteins;
- Elucidating the structure and conformational dynamics of individual proteins and large protein complexes;
- Identify binding or interaction sites when analyzing protein complexes;
- Complementarity with cryo-electron microscopy (cryo-EM), especially when crystallographic information is not available.
Sample Request
- Protein-ligand and protein samples
- Sample concentration >1ug/ul
- Sample size >10ug
- NOTE: Samples are ideally free of detergents.
In conclusion, we are confident that our hydrogen-deuterium exchange system is ideal for the high-throughput screening required by the pharmaceutical industry. Our technique is particularly suitable for analyzing the conformational dynamics of proteins in solution in small and large systems, and is complementary to many biophysical methods.
If you are interested in our services, please contact us immediately, then fill in the complete inquiry form, and we will reply to you as soon as possible.
References
- Ozohanics O, Ambrus A. Hydrogen-Deuterium Exchange Mass Spectrometry: A Novel Structural Biology Approach to Structure, Dynamics and Interactions of Proteins and Their Complexes. Life. 2020, 10(11): 286.
- Pirrone G F, et al. Applications of hydrogen/deuterium exchange MS from 2012 to 2014. Analytical chemistry. 2015, 87(1): 99-118.
