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. Author manuscript; available in PMC: 2012 Aug 24.

Published in final edited form as: Biochim Biophys Acta. 2008 Jul 23;1781(10):610–617. doi: 10.1016/j.bbalip.2008.07.002

Fig. 2 — Point mutation of individual conserved residues within D3 and D4 abolishes SMS1 activity. (A) SMS activity was performed on wild type (WT) or mutant SMS1 following immunoprecipitation from Hela cells transiently expressing the respective enzyme. The protein level of each enzyme is shown in a western blot below. (B) Wild type SMS1 is a Golgi targeted protein. Both cadherin (top) and α Mannosidase II (bottom) are depicted in green, while SMS1-Flag is depicted in red. TOPRO-3, a DNA dye is in blue. (C) Each SMS1 mutant (S283A, H285A, H328A, H332A, S273A) exhibits a WT localization pattern. α Mannosidase II is depicted in green. Flag-tagged SMS1 and mutants are depicted in red. TOPRO-3 is in blue.

Fig. 2 — Point mutation of individual conserved residues within D3 and D4 abolishes SMS1 activity. (A) SMS activity was performed on wild type (WT) or mutant SMS1 following immunoprecipitation from Hela cells transiently expressing the respective enzyme. The protein level of each enzyme is shown in a western blot below. (B) Wild type SMS1 is a Golgi targeted protein. Both cadherin (top) and α Mannosidase II (bottom) are depicted in green, while SMS1-Flag is depicted in red. TOPRO-3, a DNA dye is in blue. (C) Each SMS1 mutant (S283A, H285A, H328A, H332A, S273A) exhibits a WT localization pattern. α Mannosidase II is depicted in green. Flag-tagged SMS1 and mutants are depicted in red. TOPRO-3 is in blue.