caa a22 4500 445325054 CHVBK 20180317142717.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s10858-011-9469-5 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9469-5 Suppression of phospholipid biosynthesis by cerulenin in the condensed Single-Protein-Production (cSPP) system [Elektronische Daten] [Lili Mao, Koichi Inoue, Yisong Tao, Gaetano Montelione, Ann McDermott, Masayori Inouye] Using the single-protein-production (SPP) system, a protein of interest can be exclusively produced in high yield from its ACA-less gene in Escherichia coli expressing MazF, an ACA-specific mRNA interferase. It is thus feasible to study a membrane protein by solid-state NMR (SSNMR) directly in natural membrane fractions. In developing isotope-enrichment methods, we observed that 13C was also incorporated into phospholipids, generating spurious signals in SSNMR spectra. Notable, with the SPP system a protein can be produced in total absence of cell growth caused by antibiotics. Here, we demonstrate that cerulenin, an inhibitor of phospholipid biosynthesis, can suppress isotope incorporation in the lipids without affecting membrane protein yield in the SPP system. SSNMR analysis of ATP synthase subunit c, an E. coli inner membrane protein, produced by the SPP method using cerulenin revealed that 13C resonance signals from phospholipid were markedly reduced, while signals for the isotope-enriched protein were clearly present. Springer Science+Business Media B.V., 2011 cSPP nationallicence Membrane protein nationallicence Phospholipid biosynthesis nationallicence Cerulenin nationallicence SSNMR nationallicence Mao Lili Department of Biochemistry, Center for Advanced Biotechnology and Medicine, Robert Wood Johnson Medical School, 679 Hoes Lane, 08854, Piscataway, NJ, USA aut Inoue Koichi Department of Biochemistry, Center for Advanced Biotechnology and Medicine, Robert Wood Johnson Medical School, 679 Hoes Lane, 08854, Piscataway, NJ, USA aut Tao Yisong Department of Chemistry, Columbia University, 3000 Broadway, 10027, New York, NY, USA aut Montelione Gaetano Department of Biochemistry, Center for Advanced Biotechnology and Medicine, Robert Wood Johnson Medical School, 679 Hoes Lane, 08854, Piscataway, NJ, USA aut McDermott Ann Department of Chemistry, Columbia University, 3000 Broadway, 10027, New York, NY, USA aut Inouye Masayori Department of Biochemistry, Center for Advanced Biotechnology and Medicine, Robert Wood Johnson Medical School, 679 Hoes Lane, 08854, Piscataway, NJ, USA aut Journal of Biomolecular NMR Springer Netherlands 49/2(2011-02-01), 131-137 0925-2738 49:2<131 2011 49 10858 https://doi.org/10.1007/s10858-011-9469-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9469-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mao Lili Department of Biochemistry, Center for Advanced Biotechnology and Medicine, Robert Wood Johnson Medical School, 679 Hoes Lane, 08854, Piscataway, NJ, USA aut NATIONALLICENCE 700 1- Inoue Koichi Department of Biochemistry, Center for Advanced Biotechnology and Medicine, Robert Wood Johnson Medical School, 679 Hoes Lane, 08854, Piscataway, NJ, USA aut NATIONALLICENCE 700 1- Tao Yisong Department of Chemistry, Columbia University, 3000 Broadway, 10027, New York, NY, USA aut NATIONALLICENCE 700 1- Montelione Gaetano Department of Biochemistry, Center for Advanced Biotechnology and Medicine, Robert Wood Johnson Medical School, 679 Hoes Lane, 08854, Piscataway, NJ, USA aut NATIONALLICENCE 700 1- McDermott Ann Department of Chemistry, Columbia University, 3000 Broadway, 10027, New York, NY, USA aut NATIONALLICENCE 700 1- Inouye Masayori Department of Biochemistry, Center for Advanced Biotechnology and Medicine, Robert Wood Johnson Medical School, 679 Hoes Lane, 08854, Piscataway, NJ, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 49/2(2011-02-01), 131-137 0925-2738 49:2<131 2011 49 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer