An assay of relative cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe.
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.
De Nobel
J G
JG
Department of Molecular Cell Biology, University of Amsterdam, The Netherlands.
Klis
F M
FM
Munnik
T
T
Priem
J
J
van den Ende
H
H
D002473:Cell Wall / Q000378:metabolism / Q000648:ultrastructure; D002850:Chromatography, Gel; D003637:DEAE-Dextran / Q000494:pharmacology; D006867:Hydrolases / Q000494:pharmacology; D007716:Kluyveromyces / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D009997:Osmotic Pressure; D011107:Polylysine / Q000494:pharmacology; D016062:Porosity; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D012568:Schizosaccharomyces / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*
D011107:Polylysine; C037833:zymolyase; D003637:DEAE-Dextran; D006867:Hydrolases