Quantitative comparison of structure and dynamics of elastin following three isolation schemes by 13C solid state NMR and MALDI mass spectrometry
| Autor: | Gregory S. Boutis, Antonios Papaioannou, Basant Dhital, Marissa E Louis, Hsin-Pin Ho, Emmanuel J. Chang |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Kinetics
Biophysics Mass spectrometry Biochemistry Article Analytical Chemistry chemistry.chemical_compound Sequence Analysis Protein Magic angle spinning Animals Peptide bond Amino Acid Sequence Carbon-13 Magnetic Resonance Spectroscopy Molecular Biology Alanine Chromatography biology Temperature Elastin Protein Structure Tertiary Desmosine chemistry Solid-state nuclear magnetic resonance Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Cattle |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 1854:391-401 |
| ISSN: | 1570-9639 |
| DOI: | 10.1016/j.bbapap.2014.12.024 |
| Popis: | Methods for isolating elastin from fat, collagen, and muscle, commonly used in the design of artificial elastin based biomaterials, rely on exposing tissue to harsh pH levels and temperatures that usually denature many proteins. At present, a quantitative measurement of the modifications to elastin following isolation from other extracellular matrix constituents has not been reported. Using magic angle spinning (13)C NMR spectroscopy and relaxation methodologies, we have measured the modification in structure and dynamics following three known purification protocols. Our experimental data reveal that the (13)C spectra of the hydrated samples appear remarkably similar across the various purification methods. Subtle differences in the half maximum widths were observed in the backbone carbonyl suggesting possible structural heterogeneity across the different methods of purification. Additionally, small differences in the relative signal intensities were observed between purified samples. Lyophilizing the samples results in a reduction of backbone motion and reveals additional differences across the purification methods studied. These differences were most notable in the alanine motifs indicating possible changes in cross-linking or structural rigidity. The measured correlation times of glycine and proline moieties are observed to also vary considerably across the different purification methods, which may be related to peptide bond cleavage. Lastly, the relative concentration of desmosine cross-links in the samples quantified by MALDI mass spectrometry is reported. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect