High-level expression of the Toxoplasma gondii STT3 gene is required for suppression of the yeast STT3 gene mutation

Shams‐Eldin, Hosam; Blaschke, Thomas; Anhlan, Darisuren; Niehus, Sebastian; Müller, Jan; Azzouz, Nahid; Schwarz, Ralph Τ.

doi:10.1016/j.molbiopara.2005.04.008

Cited by 13 publications

(14 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The pooled chloroform phases (1.5 ml) were then washed eight times with ultrapure water. The methylated derivative-containing chloroform phase was finally dried under a stream of nitrogen, and the extracted products were further purified on a Sep-Pak C 18 . The Sep-Pack C 18 was sequentially conditioned with methanol (5 ml) and water (2 ϫ 5 ml).…”

Section: Methodsmentioning

confidence: 99%

“…Reduced and carboxamidomethylated Toxoplasma products were first digested with trypsin, and glycans were released from the resulting peptide/glycopeptide mixture by digestion with PNGase F. This enzyme is capable of releasing all known N-linked oligosaccharides except those with fucose attached to the 3-position of the Asn-linked GlcNAc residue. Such PNGase F-resistant oligosaccharides have been found to be sensitive to PNGase A. PNGase F-released glycans were separated from peptides and were analyzed by MALDI-TOF-MS after permethylation and purification on a Sep-Pak C 18 (Fig. 1).…”

Section: Maldi-tof-ms Of N-glycans Released From Detergent Extracts-mentioning

confidence: 99%

“…The permethylation derivatization of glycans increases the sensitivity of the detection of molecular ions. The peptide fraction was further digested with PNGase A, and the putative glycans released were analyzed by MALDI-TOF-MS after permethylation and purification on a Sep-Pak C 18 . Fig.…”

Section: Maldi-tof-ms Of N-glycans Released From Detergent Extracts-mentioning

confidence: 99%

“…The presence of N-glycoproteins is indeed still controversial (15)(16)(17)(18)(19). In general, N-glycosylation is considered a rare post-translational modification in apicomplexan parasites.…”

mentioning

confidence: 99%

See 3 more Smart Citations

Proteomics and Glycomics Analyses of N-Glycosylated Structures Involved in Toxoplasma gondii-Host Cell Interactions

Fauquenoy

Morelle

Hovasse

et al. 2008

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

The apicomplexan parasite Toxoplasma gondii recognizes, binds, and penetrates virtually any kind of mammalian cell using a repertoire of proteins released from late secretory organelles and a unique form of gliding motility (also named glideosome) that critically depends on actin filaments and myosin. How T. gondii glycosylated proteins mediate host-parasite interactions remains elusive. To date, only limited evidence is available concerning N-glycosylation in apicomplexans. Here we report comprehensive proteomics and glycomics analyses showing that several key components required for host cell-T. gondii interactions are N-glycosylated. Detailed structural characterization confirmed that N-glycans from T. gondii total protein extracts consist of oligomannosidic (Man 5-8 -(GlcNAc) 2 ) and paucimannosidic (Man 3-4 (GlcNAc) 2 ) sugars, which are rarely present on mature eukaryotic glycoproteins. In situ fluorescence using concanavalin A and Pisum sativum agglutinin predominantly stained the entire parasite body. Visualization of Toxoplasma glycoproteins purified by affinity chromatography followed by detailed proteomics and glycan analyses identified components involved in gliding motility, moving junction, and other additional functions implicated in intracellular development. Importantly tunicamycin-treated parasites were considerably reduced in motility, host cell invasion, and growth. Collectively these results indicate that N-glycosylation probably participates in modifying key proteins that are essential for host cell invasion by T. gondii. Molecular & Cellular Proteomics 7:891-910, 2008.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Maldi-tof-ms Of N-glycans Released From Detergent Extracts-mentioning

confidence: 99%

Section: Maldi-tof-ms Of N-glycans Released From Detergent Extracts-mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Proteomics and Glycomics Analyses of N-Glycosylated Structures Involved in Toxoplasma gondii-Host Cell Interactions

Fauquenoy

Morelle

Hovasse

et al. 2008

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…Although the molecular basis for the complementation of stt3 mutation by the expression of the T. gondii STT3 protein was not analyzed in detail (Shams-Eldin et al, 2005), Castro et al (2006) showed that the T. cruzi STT3 protein most likely replaced the yeast Stt3p in the complex. Our analysis revealed that such integration into the complex was not observed for the LmSTT3 proteins when expressed in yeast.…”

Section: Lmstt3d Acts Independently Of Endogenous Otase Subunits In Ymentioning

confidence: 99%

All in One:Leishmania majorSTT3 Proteins Substitute for the Whole Oligosaccharyltransferase Complex inSaccharomyces cerevisiae

Nasab¹,

Schulz²,

Gamarro

et al. 2008

MBoC

View full text Add to dashboard Cite

The transfer of lipid-linked oligosaccharide to asparagine residues of polypeptide chains is catalyzed by oligosaccharyltransferase (OTase). In most eukaryotes, OTase is a hetero-oligomeric complex composed of eight different proteins, in which the STT3 component is believed to be the catalytic subunit. In the parasitic protozoa Leishmania major, four STT3 paralogues, but no homologues to the other OTase components seem to be encoded in the genome. We expressed each of the four L. major STT3 proteins individually in Saccharomyces cerevisiae and found that three of them, LmSTT3A, LmSTT3B, and LmSTT3D, were able to complement a deletion of the yeast STT3 locus. Furthermore, LmSTT3D expression suppressed the lethal phenotype of single and double deletions in genes encoding other essential OTase subunits. LmSTT3 proteins did not incorporate into the yeast OTase complex but formed a homodimeric enzyme, capable of replacing the endogenous, multimeric enzyme of the yeast cell. Therefore, these protozoan OTases resemble the prokaryotic enzymes with respect to their architecture, but they used substrates typical for eukaryotic cells: N-X-S/T sequons in proteins and dolicholpyrophosphate-linked high mannose oligosaccharides.

show abstract

Current awareness on yeast

2006

Yeast

View full text Add to dashboard Cite

In order to keep subscribers up‐to‐date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly‐published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (3 weeks journals ‐ search completed 7th. Dec. 2004)

show abstract

High-level expression of the Toxoplasma gondii STT3 gene is required for suppression of the yeast STT3 gene mutation

Cited by 13 publications

References 21 publications

Proteomics and Glycomics Analyses of N-Glycosylated Structures Involved in Toxoplasma gondii-Host Cell Interactions

Proteomics and Glycomics Analyses of N-Glycosylated Structures Involved in Toxoplasma gondii-Host Cell Interactions

All in One:Leishmania majorSTT3 Proteins Substitute for the Whole Oligosaccharyltransferase Complex inSaccharomyces cerevisiae

Current awareness on yeast

Contact Info

Product

Resources

About