Targeting of a heterologous protein to the cell wall of <i>Saccharomyces cerevisiae</i>

Yeast - Tập 9 Số 4 - Trang 399-409 - 1993
M P Schreuder1, S.S.C. Brekelmans, H. van den Ende, Frans M. Klis
1Department of Molecular Cell Biology, University of Amsterdam, The Netherlands.

Tóm tắt

AbstractThe sexual adhesion protein of Saccharomyces cerevisiae MATα cells, α‐agglutinin, could not be extracted from the cell wall with hot sodium dodecyl sulfate (SDS), but became soluble after digestion of the cell with laminarinase. This indicates that it is intimately associated with cell wall glucan. A fusion protein was constructed consisting of the signal sequence of yeast invertase, guar α‐galactosidase, and the C‐terminal half of the α‐agglutinin. Most of the fusion protein was incorporated in the cell wall. A small amount could be extracted with SDS, but most of it could only be extracted with laminarinase. On the other hand, cells containing a construct consisting of the signal sequence of invertase and α‐galactosidase released most of the α‐galactosidase into the medium and all cell wall‐associated α‐galactosidase was released by SDS. Labelling with antibodies showed that the α‐galactosidase part of the fusion protein was exposed on the surface of the cell wall. The results demonstrate that the C‐terminal half of the α‐agglutinin contains the information needed to incorporate a protein into the cell wall.

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Tài liệu tham khảo

Ballou C. E., 1982, The Molecular Biology of the Yeast Saccharomyces. Metabolism and Gene Expression, 335

10.1146/annurev.bi.51.070182.003555

10.1002/j.1460-2075.1991.tb04984.x

10.1002/j.1460-2075.1988.tb03063.x

10.1002/yea.320060606

10.1146/annurev.bi.57.070188.001441

10.1099/00221287-98-2-315

Fleet G. H., 1981, Encyclopedia of Plant Physiology, 416

10.1021/bi00324a033

Harlow E., 1988, Antibodies, a Laboratory Manual

10.1016/0014-5793(89)81108-1

10.1016/0968-0004(90)90014-3

10.1016/0378-1119(83)90238-X

10.1126/science.6272395

10.1016/0076-6879(91)94036-C

10.1038/227680a0

10.1128/MCB.9.8.3155

10.1016/0304-4157(89)90014-2

Maniatis T., 1982, Molecular Cloning: A Laboratory Manual

10.1042/bj1350019

10.1042/bj1350031

10.1016/0378-1119(83)90126-9

10.1016/0076-6879(88)37055-2

10.1128/MCB.11.1.27

10.1007/BF00027314

10.1016/0304-4165(84)90174-0

Roy A., 1991, The AGA1 product is involved in cell surface attachment of the Saccharomyces cerevisiae cell adhesion glycoprotein a‐agglutinin, Mol. Cell. Biol., 11, 4196

10.1016/0922-338X(91)90211-X

Shibata N., 1983, Isolation of mannan‐protein complexes from viable cells of Saccharomyces cerevisiae X2180‐1A wild type and Saccharomyces cerevisiae X2180‐1A‐5 mutant strains by the action of Zymolyase 60,000, J. Bacteriol., 156, 552, 10.1128/jb.156.2.552-558.1983

10.1002/yea.320090102

10.1073/pnas.76.9.4350

10.1016/S0021-9258(17)43167-X

10.1016/0167-4838(90)90237-A

10.1016/S0021-9258(18)98888-5

10.1099/00221287-130-6-1419

10.1002/yea.320070707

10.1002/j.1460-2075.1988.tb02966.x

Wojciechowicz D.(1991). The immunological and molecular characterization of α‐agglutinin fromSaccharomyces cerevisiae. PhD thesis City University of New York New York USA.

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Yeast

Tập 9 Số 4

399-409

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