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Part II: defining and quantifying individual and co-cultured intracellular proteomes of two thermophilic microorganisms by GeLC-MS2 and spectral counting

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An Erratum to this article was published on 13 August 2010

Abstract

Probing the intracellular proteome of Thermotoga maritima and Caldicellulosiruptor saccharolyticus in pure and co-culture affords a global investigation into the machinery and mechanisms enduring inside the bacterial thermophilic cell at the time of harvest. The second of a two part study, employing GeLC-MS2 a variety of proteins were confidently identified with <1% false discovery rate, and spectral counts for label-free relative quantification afforded indication of the dynamic proteome as a function of environmental stimuli. Almost 25% of the T. maritima proteome and 10% of the C. saccharolyticus proteome were identified. Through comparison of growth temperatures for T. maritima, a protein associated with chemotaxis was uniquely present in the sample cultivated at the non-optimal growth temperature. It is suspected that movement was induced due to the non-optimal condition as the organism may need to migrate in the culture to locate more nutrients. The inventory of C. saccharolyticus proteins identified in these studies and attributed to spectral counting, demonstrated that two CRISPR-associated proteins had increased expression in the pure culture versus the co-culture. Further focusing on this relationship, a C. saccharolyticus phage-shock protein was identified in the co-culture expanding a scenario that the co-culture had decreased antiviral resistance and accordingly an infection-related protein was present. Alterations in growth conditions of these bacterial thermophilic microorganisms offer a glimpse into the intricacy of microbial behavior and interaction.

A summary comparing the number of confidently identified proteins from the cell lysis of the thermophilic bacteria T. maritima and C. saccharolyticus in mono- and co-culture. The expression profile of identified proteins provides insight into microbial interaction and behavior.

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Acknowledgments

We would like to acknowledge the financial support of the National Institutes of Health (Grant 5T32GM00-8776-08), which supports GLA in the North Carolina State University Molecular Biotechnology Training Program, and the W. M. Keck Foundation. RMK acknowledges support from the US National Science Foundation (CBT0617272) and the Bioenergy Science Center (BESC), a U.S. DOE Bioenergy Research Center supported by the Office of Biological and Environmental Research. DLL acknowledges support from a US Department of Education GAANN Fellowship.

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Authors and Affiliations

  1. Chemistry, North Carolina State University, Raleigh, NC, 27695, USA

    David Muddiman & Genna Andrews

  2. Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, 27695, USA

    Derrick Lewis, Jaspreet Notey & Robert Kelly

  3. W.M. Keck FT-ICR Mass Spectrometry Laboratory, North Carolina State University, Raleigh, NC, 27695, USA

    David Muddiman

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  1. David Muddiman
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Correspondence to David Muddiman.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s00216-010-4050-8

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Muddiman, D., Andrews, G., Lewis, D. et al. Part II: defining and quantifying individual and co-cultured intracellular proteomes of two thermophilic microorganisms by GeLC-MS2 and spectral counting. Anal Bioanal Chem 398, 391–404 (2010). https://doi.org/10.1007/s00216-010-3929-8

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