Document Detail


A simple and specific procedure to permeabilize the plasma membrane of Schizosaccharomyces pombe.
MedLine Citation:
PMID:  19734666     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cu(2+)-treatment is a useful technique in selectively permeabilizing the fungal plasma membrane. We describe herein a practical application with Schizosaccharomyces pombe. Incubation of cells with 0.5 mM CuCl(2) at 30 degrees C for 20 min induced efficient leakage of cytosolic constituents. The kinetic characteristics of the calcium and amino acid flux from Cu(2+)-treated S. pombe cells suggested that the Cu(2+) treatment permeabilized the plasma membrane without loss of vacuolar function. As a further application of the method, the amino acid contents of Cu(2+)-treated and untreated cells were also determined. The amino acid pool of Cu(2+)-treated wild-type cells was enriched in basic amino acids but not in acidic amino acids, as is characteristic of the vacuolar amino acid pool of fungi, including Saccharomyces cerevisiae and Neurosporra crassa. The amino acid pool of the S. pombe V-ATPase mutant vma1Delta was also successfully determined. We conclude that the vacuolar amino acid pool of S. pombe can be measured using Cu(2+)-treated cells. The method is simple, inexpensive, and rapid relative to the isolation of vacuolar vesicles, making it useful in estimating vacuolar pools and transport across the vacuolar membrane.
Authors:
Soracom Chardwiriyapreecha; Kana Hondo; Hiroko Inada; Thippayarat Chahomchuen; Takayuki Sekito; Tomoko Iwaki; Yoshimi Kakinuma
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-09-07
Journal Detail:
Title:  Bioscience, biotechnology, and biochemistry     Volume:  73     ISSN:  1347-6947     ISO Abbreviation:  Biosci. Biotechnol. Biochem.     Publication Date:  2009 Sep 
Date Detail:
Created Date:  2009-09-24     Completed Date:  2009-12-31     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9205717     Medline TA:  Biosci Biotechnol Biochem     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  2090-5     Citation Subset:  IM    
Affiliation:
Laboratory of Molecular Physiology and Genetics, Faculty of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama 790-8566, Japan.
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / metabolism
Calcium / metabolism
Cell Membrane Permeability / drug effects*
Copper / pharmacology*
Kinetics
Schizosaccharomyces / cytology*,  enzymology,  metabolism
Vacuolar Proton-Translocating ATPases / metabolism
Chemical
Reg. No./Substance:
0/Amino Acids; 7440-50-8/Copper; 7440-70-2/Calcium; 7447-39-4/cupric chloride; EC 3.6.1.-/Vacuolar Proton-Translocating ATPases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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