Screening for glycosylphosphatidylinositol (GPI)-dependent cell wall proteins in Saccharomyces cerevisiae.

摘要: Open reading frames in the genome of Saccharomyces cerevisiae were screened for potential glycosylphosphatidylinositol (GPI)-attached proteins. The identification putative GPI-attached proteins was based on three criteria: presence a GPI-attachment signal sequence, sequence secretion and serine- or threonine-rich sequence. In all, 53 ORFs met these criteria 38 further analyzed as follows. encoding 40 C-terminal amino acids each fused with structural gene reporter protein consisting signal, α-galactosidase hemagglutinin (HA) epitope, examined ability to become incorporated into cell wall. On this basis, 14 fusion classified GPI-dependent wall because cells expressing proteins: (i) had high levels activity their surface; (ii) released significant amounts from membrane treatment phosphatidylinositol-specific phospholipase C (PI-PLC); (iii) following laminarinase. Of identified proteins, 12 novel adjacent Amino acid alignment sequences ORFs, together those known reveals some similarities among them.