AN ASSAY OF RELATIVE CELL-WALL POROSITY IN SACCHAROMYCES-CEREVISIAE, KLUYVEROMYCES-LACTIS AND SCHIZOSACCHAROMYCES-POMBE

被引:124
作者
DENOBEL, JG
KLIS, FM
MUNNIK, T
PRIEM, J
VANDENENDE, H
机构
[1] Department of Molecular Cell Biology, Biotechnology Center, University of Amsterdam, Amsterdam, 1098 SM
关键词
Cell wall porosity; cell wall structure; permeability; polycation assay;
D O I
10.1002/yea.320060605
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have developed a new assay to determine relative cell wall porosity in yeasts, which is based on polycation‐induced leakage of UV‐absorbing compounds. Polycations with a small hydrodynamic radius as measured by gel filtration (poly‐L‐lysine) caused cell leakage independent of cell wall porosity whereas polycations with a large hydrodynamic radius (DEAE‐dextrans) caused only limited cell leakage due to limited passage through the cell wall. This allowed the ratio between DEAE‐dextran‐ and poly‐L‐lysine‐induced cell leakage to be used as a measure of cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe. Using this assay, we found that the composition of the growth medium affected cell wall porosity in S. cerevisiae. In addition, we could show that cell wall porosity is limited by the number of disulphide bridges in the wall and is dependent on cell turgor. It is argued that earlier methods to estimate cell wall porosity in S. cerevisiae resulted in large underestimations. Copyright © 1990 John Wiley & Sons Ltd.
引用
收藏
页码:483 / 490
页数:8
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