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Specific labelling of cell wall proteins by biotinylation. Identification of four covalently linked O-mannosylated proteins of Saccharomyces cerevisiae.
Yeast. 1997 Sep 30; 13(12):1145-54.Y

Abstract

Intact Saccharomyces cerevisiae cells were biotinylated with the non-permeable sulfosuccinimidyl-6-(biotinamido) hexanoate reagent. Twenty specifically labelled cell wall proteins would be extracted and visualized on SDS gels via streptavidin/horseradish peroxidase. Nine cell wall proteins were released by SDS extraction under reducing conditions and were designated Scw1-9p for (soluble cell wall proteins); five proteins were released from SDS-extracted cell walls by laminarinase (Ccw1-5p for covalently linked cell wall proteins) and six with mild (30 mM-NaOH, 4 degrees C, 14 h) alkali treatment (Ccw6-11p). N-terminal sequences of the Ccw proteins 6, 7, 8 and 11 showed that these cell wall proteins are members of the PIR gene family (predicted proteins with internal repeats), CCW6 being identical to PIR1 and CCW8 to PIR3. Single gene disruptions of all four genes did not yield a phenotype. In the CCW11 disruption the Ccw11p as well as the laminarinase-extracted Ccw5 protein was missing. The new cell wall proteins are O-mannosylated, contain a Kex2 processing site, but no C-terminal GPI anchor sequence.

Authors+Show Affiliations

Mrsă V
Lehrstuhl für Zellbiologie und Pflanzenphysiologie, Universität Regensburg, Germany.
Seidl T
No affiliation info available
Gentzsch M
No affiliation info available
Tanner W
No affiliation info available

MeSH

Amino Acid SequenceBiotinCell WallFungal ProteinsGlycosylationMannoseMolecular Sequence DataSaccharomyces cerevisiae

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9301021

Citation

Mrsă, V, et al. "Specific Labelling of Cell Wall Proteins By Biotinylation. Identification of Four Covalently Linked O-mannosylated Proteins of Saccharomyces Cerevisiae." Yeast (Chichester, England), vol. 13, no. 12, 1997, pp. 1145-54.
Mrsă V, Seidl T, Gentzsch M, et al. Specific labelling of cell wall proteins by biotinylation. Identification of four covalently linked O-mannosylated proteins of Saccharomyces cerevisiae. Yeast. 1997;13(12):1145-54.
Mrsă, V., Seidl, T., Gentzsch, M., & Tanner, W. (1997). Specific labelling of cell wall proteins by biotinylation. Identification of four covalently linked O-mannosylated proteins of Saccharomyces cerevisiae. Yeast (Chichester, England), 13(12), 1145-54.
Mrsă V, et al. Specific Labelling of Cell Wall Proteins By Biotinylation. Identification of Four Covalently Linked O-mannosylated Proteins of Saccharomyces Cerevisiae. Yeast. 1997 Sep 30;13(12):1145-54. PubMed PMID: 9301021.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Specific labelling of cell wall proteins by biotinylation. Identification of four covalently linked O-mannosylated proteins of Saccharomyces cerevisiae. AU - Mrsă,V, AU - Seidl,T, AU - Gentzsch,M, AU - Tanner,W, PY - 1997/9/25/pubmed PY - 2000/6/20/medline PY - 1997/9/25/entrez SP - 1145 EP - 54 JF - Yeast (Chichester, England) JO - Yeast VL - 13 IS - 12 N2 - Intact Saccharomyces cerevisiae cells were biotinylated with the non-permeable sulfosuccinimidyl-6-(biotinamido) hexanoate reagent. Twenty specifically labelled cell wall proteins would be extracted and visualized on SDS gels via streptavidin/horseradish peroxidase. Nine cell wall proteins were released by SDS extraction under reducing conditions and were designated Scw1-9p for (soluble cell wall proteins); five proteins were released from SDS-extracted cell walls by laminarinase (Ccw1-5p for covalently linked cell wall proteins) and six with mild (30 mM-NaOH, 4 degrees C, 14 h) alkali treatment (Ccw6-11p). N-terminal sequences of the Ccw proteins 6, 7, 8 and 11 showed that these cell wall proteins are members of the PIR gene family (predicted proteins with internal repeats), CCW6 being identical to PIR1 and CCW8 to PIR3. Single gene disruptions of all four genes did not yield a phenotype. In the CCW11 disruption the Ccw11p as well as the laminarinase-extracted Ccw5 protein was missing. The new cell wall proteins are O-mannosylated, contain a Kex2 processing site, but no C-terminal GPI anchor sequence. SN - 0749-503X UR - https://www.unboundmedicine.com/medline/citation/9301021/Specific_labelling_of_cell_wall_proteins_by_biotinylation__Identification_of_four_covalently_linked_O_mannosylated_proteins_of_Saccharomyces_cerevisiae_ L2 - https://doi.org/10.1002/(SICI)1097-0061(19970930)13:12<1145::AID-YEA163>3.0.CO;2-Y DB - PRIME DP - Unbound Medicine ER -