Title: Localization and targeting of the Saccharomyces cerevisiae Kre2p/Mnt1p alpha 1,2-mannosyltransferase to a medial-Golgi compartment Document date: 1995_11_2
ID: q1jx0n0l_13
Snippet: Analysis of [35S]methionine-labeled proteins was as described (Cooper and Bussey, 1989) . Briefly, cells were grown to an OD600 of 0.4-0.45 in selective medium (YNB) and then labeled with Trans35S (100 ~Ci; ICN Biochemicals Inc., Montr6al, Canada) for 10 min at 30°C. Yeast cells treated with tunicamycin (10 ~g m1-1) were preincubated before radiolabeling for 30 min at 30°C. Tunicamycin is a hydrophobic analogue of UDP-N-acetylglucosamine that b.....
Document: Analysis of [35S]methionine-labeled proteins was as described (Cooper and Bussey, 1989) . Briefly, cells were grown to an OD600 of 0.4-0.45 in selective medium (YNB) and then labeled with Trans35S (100 ~Ci; ICN Biochemicals Inc., Montr6al, Canada) for 10 min at 30°C. Yeast cells treated with tunicamycin (10 ~g m1-1) were preincubated before radiolabeling for 30 min at 30°C. Tunicamycin is a hydrophobic analogue of UDP-N-acetylglucosamine that blocks the addition of N-acetylglucosamine to dolichol phosphate, the first step in N-linked oligosaccharide formation. Chase conditions were achieved by addition of L-methionine and L-cysteine to a final concentration of i mM.
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