PMID: 375979May 17, 1979Paper

Yeast plasma membrane ghosts. An analysis of proteins by two-dimensional gel electrophoresis

Biochimica Et Biophysica Acta
H BusseyG H Rank

Abstract

We have examined yeast cell ghost preparations to assess their value in obtaining plasma membrane proteins. Ghosts prepared by two methods involving stabilization of spheroplast envelopes had similar protein patterns by two-dimensional gel electrophoresis, and approximately 200 proteins were resolved. Spheroplasts were lactoperoxidase iodinated, and recovery of label in ghost preparations was greater than 60%. Spheroplasts appeared to be impermeable to the lactoperoxidase reagents as judged by an examination of two-dimensional gel electrophoretic patterns of ghost proteins that had been iodinated in spheroplasts or in unsealed ghosts. Spheroplasts were also impermeable to pronase proteases. Surface iodination and surface proteolysis allowed us to identify exposed ghost proteins; the major ghost glycoprotein was exposed in spheroplasts. Two-dimensional patterns of ghost proteins were not heavily contaminated (less than or equal to 25% of all proteins) by proteins present in soluble or promitochondrial fractions, and estimates of surface label and total cell protein recovery suggested that the ghost fraction represents a cell envelope enrichment of 8--10 fold over whole cells. Resolution of ghost proteins by two-dimensional gel e...Continue Reading

References

Apr 1, 1975·Analytical Biochemistry·R B Needleman, A Tzagoloff
Mar 21, 1978·Biochimica Et Biophysica Acta·E SantosR Sentandreu
Feb 10, 1976·Biochemistry·G F Ames, K Nikaido
Oct 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·A DuránE Cabib
Nov 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·R O Hynes
Oct 23, 1974·Biochemical and Biophysical Research Communications·J F LenneyJ Meyer
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Nov 1, 1953·Proceedings of the Society for Experimental Biology and Medicine·J BADINM SCHUBERT
Feb 1, 1958·Biochimica Et Biophysica Acta·H HALVORSON

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Citations

Feb 1, 1985·Journal of Bioenergetics and Biomembranes·E Marrè, A Ballarin-Denti
Jul 1, 1996·Journal of Photochemistry and Photobiology. B, Biology· Fraikin GYaA B Rubin
Sep 12, 1979·Biochemical and Biophysical Research Communications·D T RogersH Bussey
Mar 8, 1982·Biochimica Et Biophysica Acta·E SantosR Sentandreu
Oct 1, 1983·Journal of Bacteriology·J Tschopp, R Schekman
Dec 30, 1981·Biochimica Et Biophysica Acta·A Goffeau, C W Slayman

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