Procollagen galactosyltransferase 1

GLT25D1/COLGALT1 was produced recombinantly as a N-terminal 8xHis-SUMO fusion in E. coli. After initial affinity chromatography, the sample was subject to tag removal using His-tagged SUMO protease followed by reverse affinity chromatography and dialysis. The sample was then subject to anion exchange chromatography for further polishing, followed by gel filtration into a Superdex 200 Increase 10/300 GL (Cytiva), equilibrated with 25 mM HEPES/NaOH, 100 mM NaCl, pH 8.0. The sample was concentrated to 4 mg/mL in a 30 kDa MWCO Vivaspin Turbo centrifugal filter (Sartorius) and used for SEC-SAXS analysis