Extended Data Fig. 1: Biochemical and functional characterisation of ScALG3, HsALG9 and GgALG12.
From: Structures of ALG3/9/12 reveal the assembly logic of the N-glycan oligomannose core
a, Size exclusion chromatography of ScALG3, HsALG9 and GgALG12 solubilized in LMNG-CHS detergent. b, SDS-PAGE analysis of ScALG3, HsALG9 and GgALG12. Representative gel shown from multiple (n > 10) independent purifications of ALG3, ALG9 and ALG12. Molecular weight markers are indicated in kDa. c-e, Assays with WT and catalytic mutants of c, ScALG3, d, HsALG9 and e, GgALG12 in LMNG-CHS. The reaction products were transferred to a fluorescent peptide (TAMRA-YANATS-NH2) using TbSTT3B and separated by tricine gel electrophoresis. The reaction times for each sample are indicated above the lanes and are coloured red for inactive, green for active and black for partially active. Lanes with glycopeptide cartoons over them serve as glycopeptide standards for the indicated glycan. Activity assays for each construct were performed at least twice with similar results.
