Literature DB >> 23742073

Quantitation of recombinant protein in whole cells and cell extracts via solid-state NMR spectroscopy.

Erica P Vogel1, David P Weliky.   

Abstract

Recombinant proteins (RPs) are commonly expressed in bacteria followed by solubilization and chromatography. Purified RP yield can be diminished by losses at any step with very different changes in methods that can improve the yield. Time and labor can therefore be saved by first identifying the specific reason for the low yield. This study describes a new solid-state nuclear magnetic resonance approach to RP quantitation in whole cells or cell extracts without solubilization or purification. The method is straightforward and inexpensive and requires only ∼50 mL culture and a low-field spectrometer.

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Year:  2013        PMID: 23742073      PMCID: PMC3734946          DOI: 10.1021/bi4007034

Source DB:  PubMed          Journal:  Biochemistry        ISSN: 0006-2960            Impact factor:   3.162


  21 in total

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Journal:  Proc Natl Acad Sci U S A       Date:  1999-08-03       Impact factor: 11.205

2.  Expression, purification, and PC1-mediated processing of (H10D, P28K, and K29P)-human proinsulin.

Authors:  Robert B Mackin; Meredith H Choquette
Journal:  Protein Expr Purif       Date:  2003-02       Impact factor: 1.650

3.  In-cell solid-state NMR as a tool to study proteins in large complexes.

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Journal:  Chembiochem       Date:  2012-02-01       Impact factor: 3.164

4.  Fast quantification of recombinant protein inclusion bodies within intact cells by FT-IR spectroscopy.

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Journal:  Biotechnol Prog       Date:  2007-05-11

5.  Solid-state NMR structural measurements on the membrane-associated influenza fusion protein ectodomain.

Authors:  Jaime Curtis-Fisk; Casey Preston; Zhaoxiong Zheng; R Mark Worden; David P Weliky
Journal:  J Am Chem Soc       Date:  2007-08-25       Impact factor: 15.419

6.  Solid-state REDOR NMR distance measurements at the ligand site of a bacterial chemotaxis membrane receptor.

Authors:  J Wang; Y S Balazs; L K Thompson
Journal:  Biochemistry       Date:  1997-02-18       Impact factor: 3.162

7.  Nutrient-dependent structural changes in S. aureus peptidoglycan revealed by solid-state NMR spectroscopy.

Authors:  Xiaoxue Zhou; Lynette Cegelski
Journal:  Biochemistry       Date:  2012-10-02       Impact factor: 3.162

8.  The crystal structure of the SIV gp41 ectodomain at 1.47 A resolution.

Authors:  Z N Yang; T C Mueser; J Kaufman; S J Stahl; P T Wingfield; C C Hyde
Journal:  J Struct Biol       Date:  1999-06-15       Impact factor: 2.867

9.  Isotopically labeled expression in E. coli, purification, and refolding of the full ectodomain of the influenza virus membrane fusion protein.

Authors:  Jaime Curtis-Fisk; Ryan M Spencer; David P Weliky
Journal:  Protein Expr Purif       Date:  2008-06-28       Impact factor: 1.650

10.  Oritavancin exhibits dual mode of action to inhibit cell-wall biosynthesis in Staphylococcus aureus.

Authors:  Sung Joon Kim; Lynette Cegelski; Dirk Stueber; Manmilan Singh; Evelyne Dietrich; Kelly S E Tanaka; Thomas R Parr; Adel Rafai Far; Jacob Schaefer
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  7 in total

1.  A new understanding of antibiotic action via solid-state NMR of cells with uniform isotopic labeling.

Authors:  David P Weliky
Journal:  Biophys J       Date:  2015-03-24       Impact factor: 4.033

2.  Full-length trimeric influenza virus hemagglutinin II membrane fusion protein and shorter constructs lacking the fusion peptide or transmembrane domain: Hyperthermostability of the full-length protein and the soluble ectodomain and fusion peptide make significant contributions to fusion of membrane vesicles.

Authors:  Punsisi U Ratnayake; E A Prabodha Ekanayaka; Sweta S Komanduru; David P Weliky
Journal:  Protein Expr Purif       Date:  2015-08-19       Impact factor: 1.650

Review 3.  Bacterial cell wall composition and the influence of antibiotics by cell-wall and whole-cell NMR.

Authors:  Joseph A H Romaniuk; Lynette Cegelski
Journal:  Philos Trans R Soc Lond B Biol Sci       Date:  2015-10-05       Impact factor: 6.237

Review 4.  Membrane protein structure from rotational diffusion.

Authors:  Bibhuti B Das; Sang Ho Park; Stanley J Opella
Journal:  Biochim Biophys Acta       Date:  2014-04-18

Review 5.  NMR of Macromolecular Assemblies and Machines at 1 GHz and Beyond: New Transformative Opportunities for Molecular Structural Biology.

Authors:  Caitlin M Quinn; Mingzhang Wang; Tatyana Polenova
Journal:  Methods Mol Biol       Date:  2018

6.  Efficient Fusion at Neutral pH by Human Immunodeficiency Virus gp41 Trimers Containing the Fusion Peptide and Transmembrane Domains.

Authors:  S Liang; P U Ratnayake; C Keinath; L Jia; R Wolfe; A Ranaweera; D P Weliky
Journal:  Biochemistry       Date:  2018-02-06       Impact factor: 3.162

7.  Folded monomers and hexamers of the ectodomain of the HIV gp41 membrane fusion protein: potential roles in fusion and synergy between the fusion peptide, hairpin, and membrane-proximal external region.

Authors:  Koyeli Banerjee; David P Weliky
Journal:  Biochemistry       Date:  2014-11-14       Impact factor: 3.162

  7 in total

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