To-date, the complete structural characterization of a protein by solution or solid-state NMR spectroscopy has typically implemented a series of experiments to assign chemical shifts (and, hence, secondary structure) to each amino acid in the sequence. Although feasible for a growing number of proteins and peptides, such is not the case for elastin. Elastin is assembled from its soluble precursor, tropoelastin, which is a high-molecular weight protein with numerous repeating sequences. Indeed, the polymeric character of both tropoelastin and elastin required a different approach, which is described in this talk. Our strategy combines enrichment of the key amino acid types in elastin with NMR experiments that target specific pairs of amino acids. Specifically, the crosslinking and hydrophobic domains are distinguished by the abundant Ala-neighboring-Ala and Gly-neighboring-Gly, respectively. With multiple populations of Gly and Pro, selection via 13C-15N and 13C-13C dipolar and/or J couplings was employed to unambiguously identify the targeted 2-aa sequences. These results and their implications for the understanding of elastin’s elasticity will be discussed.

Targeting repeating subunits in elastin with strategic enrichment and solid-state NMR spectroscopy

255th National Meeting (2018)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.