Document Detail


Analysis of chloroquine resistance transporter (CRT) isoforms and orthologues in S. cerevisiae yeast.
MedLine Citation:
PMID:  21744797     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Previous work from our laboratory optimized MeOH-inducible expression of the P. falciparum malarial parasite transporter PfCRT in P. pastoris yeast. These strains are useful for many experiments but do not allow for inducible protein expression under ambient growth conditions. We have therefore optimized galactose-inducible expression of PfCRT in S. cerevisiae yeast. We find that expression of PfCRT confers CQ hypersensitivity to growing yeast and that this is due to plasma membrane localization of the transporter. We use quantitative analyses of growth rates to compare hypersensitivity for yeast expressing various PfCRT isoforms. We also report successful high level inducible expression of the P. vivax orthologue, PvCRT, and compare CQ hypersensitivity for PvCRT vs PfCRT expressing yeast. We test the hypothesis that hypersensitivity is due to increased transport of CQ into yeast expressing the transporters via direct (3)H-CQ transport experiments and analyze the effect that membrane potential has on transport. The data suggest important new tools for rapid functional screening of PfCRT and PvCRT isoforms and provide further evidence for a model wherein membrane potential promotes charged CQ transport by PfCRT. Data also support our previous conclusion that wild type PfCRT is capable of CQ transport and provide a basis for understanding the lack of correspondence between PvCRT mutations and resistance to CQ in the important malarial parasite P. vivax.
Authors:
Nicholas K Baro; Chaya Pooput; Paul D Roepe
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, N.I.H., Extramural     Date:  2011-07-19
Journal Detail:
Title:  Biochemistry     Volume:  50     ISSN:  1520-4995     ISO Abbreviation:  Biochemistry     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-08-02     Completed Date:  2011-11-21     Revised Date:  2014-09-22    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  United States    
Other Details:
Languages:  eng     Pagination:  6701-10     Citation Subset:  IM    
Copyright Information:
© 2011 American Chemical Society
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MeSH Terms
Descriptor/Qualifier:
Animals
Antimalarials / chemistry,  pharmacology
Cell Membrane / chemistry,  genetics,  physiology
Chloroquine / chemistry*,  pharmacology
Galactose / pharmacology
Gene Expression Regulation, Fungal* / drug effects
Membrane Potentials / drug effects,  genetics
Membrane Transport Proteins / biosynthesis,  chemistry*,  genetics
Plasmodium falciparum / chemistry*,  genetics,  growth & development
Plasmodium vivax / chemistry*,  genetics,  growth & development
Protein Isoforms / biosynthesis,  chemistry,  genetics
Protozoan Proteins / biosynthesis,  chemistry*,  genetics
Saccharomyces cerevisiae / drug effects,  genetics*,  growth & development
Grant Support
ID/Acronym/Agency:
AI056312/AI/NIAID NIH HHS; AI090832/AI/NIAID NIH HHS; R01 AI056312/AI/NIAID NIH HHS; R01 AI056312-07/AI/NIAID NIH HHS; R56 AI090832/AI/NIAID NIH HHS; R56 AI090832-01/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Antimalarials; 0/Membrane Transport Proteins; 0/PfCRT protein, Plasmodium falciparum; 0/Protein Isoforms; 0/Protozoan Proteins; 886U3H6UFF/Chloroquine; X2RN3Q8DNE/Galactose
Comments/Corrections

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


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