Sharma, Alok K.Krieger, TobiasRigby, Alan C.Zelikovic, IsraelAlper, Seth2017-12-052016Sharma, Alok K., Tobias Krieger, Alan C. Rigby, Israel Zelikovic, and Seth L. Alper. 2016. “Human SLC26A4/Pendrin STAS domain is a nucleotide-binding protein: Refolding and characterization for structural studies.” Biochemistry and Biophysics Reports 8 (1): 184-191. doi:10.1016/j.bbrep.2016.08.022. http://dx.doi.org/10.1016/j.bbrep.2016.08.022.http://nrs.harvard.edu/urn-3:HUL.InstRepos:34491975Mutations in the human SLC26A4/Pendrin polypeptide (hPDS) cause Pendred Syndrome /DFNB4, syndromic deafness with enlargement of the vestibular aqueduct and low-penetrance goiter. Here we present data on cloning, protein overexpression and purification, refolding, and biophysical characterization of the recombinant hPDS STAS domain lacking its intrinsic variable sequence (STAS-ΔIVS). We report a reproducible protein refolding protocol enabling milligram scale expression and purification of uniformly 15N- and 13C/15N-enriched hPDS STAS-ΔIVS domain suitable for structural characterization by solution NMR. Circular dichroism, one-dimensional 1H, two-dimensional 1H–15N HSQC, and 1H–13C HSQC NMR spectra confirmed the well-folded state of purified hPDS STAS-ΔIVS in solution. Heteronuclear NMR chemical shift perturbation of select STAS-ΔIVS residues by GDP was observed at fast-to-intermediate NMR time scales. Intrinsic tryptophan fluorescence quench experiments demonstrated GDP binding to hPDS STAS-ΔIVS with Kd of 178 μM. These results are useful for structure/function characterization of hPDS STAS, the cytoplasmic subdomain of the congenital deafness protein, pendrin, as well as for studies of other mammalian STAS domains.en-UShPDS, Human pendrin polypeptideIVS, Intrinsic variable sequenceHSQC, Heteronuclear single quantum correlation spectroscopyCD, Circular dichroismMS, Mass spectroscopySLC26A4STAS domainIVSProtein refoldingNMRFluorescenceJournal Article2017-12-0510.1016/j.bbrep.2016.08.022