High-yield recombinant bacterial expression of ¹³ C-, ¹⁵ N-labeled, serine-16 phosphorylated, murine amelogenin using a modified third generation genetic code expansion protocol.

ABSTRACT: Amelogenin constitutes ~90% of the enamel matrix in the secretory stage of amelogenesis, a still poorly understood process that results in the formation of the hardest and most mineralized tissue in vertebrates-enamel. Most biophysical research with amelogenin uses recombinant protein expressed in Escherichia coli. In addition to providing copious amounts of protein, recombinant expression allows ¹³ C- and ¹⁵ N-labeling for detailed structural studies using NMR spectroscopy. However, native amelogenin is phosphorylated at one position, Ser-16 in murine amelogenin, and there is mounting evidence that Ser-16 phosphorylation is important. Using a modified genetic code expansion protocol we have expressed and purified uniformly ¹³ C-, ¹⁵ N-labeled mu

SUBMITTER: Buchko GW 

PROVIDER: S-EPMC9850436 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature