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Altered Escherichia coli membrane protein assembly machinery allows proper membrane assembly of eukaryotic protein vitamin K epoxide reductase.
Hatahet, Feras; Blazyk, Jessica L; Martineau, Eugenie; Mandela, Eric; Zhao, Yongxin; Campbell, Robert E; Beckwith, Jonathan; Boyd, Dana.
Afiliación
  • Hatahet F; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115;
  • Blazyk JL; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115;
  • Martineau E; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115;
  • Mandela E; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115;
  • Zhao Y; Department of Chemistry, University of Alberta, Edmonton, AB T6G 2R3, Canada.
  • Campbell RE; Department of Chemistry, University of Alberta, Edmonton, AB T6G 2R3, Canada.
  • Beckwith J; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115; jon_beckwith@hms.harvard.edu.
  • Boyd D; Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115;
Proc Natl Acad Sci U S A ; 112(49): 15184-9, 2015 Dec 08.
Article en En | MEDLINE | ID: mdl-26598701
Functional overexpression of polytopic membrane proteins, particularly when in a foreign host, is often a challenging task. Factors that negatively affect such processes are poorly understood. Using the mammalian membrane protein vitamin K epoxide reductase (VKORc1) as a reporter, we describe a genetic selection approach allowing the isolation of Escherichia coli mutants capable of functionally expressing this blood-coagulation enzyme. The isolated mutants map to components of membrane protein assembly and quality control proteins YidC and HslV. We show that changes in the VKORc1 sequence and in the YidC hydrophilic groove along with the inactivation of HslV promote VKORc1 activity and dramatically increase its expression level. We hypothesize that such changes correct for mismatches in the membrane topogenic signals between E. coli and eukaryotic cells guiding proper membrane integration. Furthermore, the obtained mutants allow the study of VKORc1 reaction mechanisms, inhibition by warfarin, and the high-throughput screening for potential anticoagulants.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Escherichia coli / Escherichia coli / Vitamina K Epóxido Reductasas / Proteínas de la Membrana Límite: Animals Idioma: En Revista: Proc Natl Acad Sci U S A Año: 2015 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Escherichia coli / Escherichia coli / Vitamina K Epóxido Reductasas / Proteínas de la Membrana Límite: Animals Idioma: En Revista: Proc Natl Acad Sci U S A Año: 2015 Tipo del documento: Article Pais de publicación: Estados Unidos