Figure 3

Glycosylation defects in C. albicans MNN1 family null mutants. The extent of N-glycosylation was determined by activity staining of Hex1 (β-N-acetylhexosaminidase) after protein samples were separated by non-denaturing gel electrophoresis ( A ). The continuous and dotted lines indicate the normal and decreased electrophoretic mobility of Hex1 respectively. The degree of N-glycosylation modification was also determined by western blot analysis of Hex1-V5-6× His tagged strains ( B ). Hex1-V5-6× His is apparent in both an unmodified (67 kDa) and a heavily glycosylated (~125 kDa) form. Acid-labile phosphomannan ( C ) and β-eliminated O-mannan ( D ) were released from [2-³H] mannose-labelled cells and separated by TLC. Samples are as follows: lane 1, wild type; lane 2, mnn1∆; lane 3, mnn12∆; lane 4, mnn13∆; lane 5, mnn14∆; lane 6, mnn15∆; lane 7, mnn16∆; lane 8, och1∆; lane 9, pmr1∆; lane 10, mnt1∆-mnt2∆.