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Association of the Rv0679c protein with lipids and carbohydrates in Mycobacterium tuberculosis/Mycobacterium bovis BCG

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Abstract

The Rv0679c gene in Mycobacterium tuberculosis H37Rv encodes a protein with a predicted molecular mass of 16,586 Da consisting of 165 amino acids which contains a putative N-terminal signal sequence and a consensus lipoprotein-processing motif. Globomycin treatment, Triton X-114 separation and mass spectrometry analyses clarified a property of the Rv0679c protein as a lipoprotein. In addition, trifluoromethanesulphonic acid treatment of the lysate revealed an association of the recombinant Rv0679c protein with carbohydrates. The Rv0679c protein homolog of Mycobacterium bovis BCG was also expressed as the protein associated with lipids and carbohydrates. In Western blot analysis, each of the protein homolog and Lipoarabinomannan (LAM) was detected as a similar pattern by anti-Rv0679c and anti-LAM antibodies, respectively. Interestingly, the Rv0679c protein was detected in commercially available LAM purified from M. tuberculosis. Inhibition assay of LAM synthesis in M. bovis BCG by ethambutol showed an altered migration pattern of the Rv0679c protein to low molecular mass similar to that of LAM. The results suggest that the Rv0679c protein exists as a tight complex with LAM in M. tuberculosis/M. bovis BCG.

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Abbreviations

LAM:

Lipoarabinomannan

LM:

Lipomannan

PIM:

Phosphatidyl-myo-inositol mannosides

TFMS:

Trifluoromethanesulfonic acid

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Acknowledgments

We are grateful to Sankyo Chemical Co., Ltd., Tokyo, Japan for the generous gift of globomycin.

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  1. Division of Bacteriology, Department of Microbiology and Immunology, Faculty of Medicine, Tottori University, 86 Nishi-cho, Yonago, Tottori, 683-8503, Japan

    Takashi Matsuba & Yoshinori Tanaka

  2. Department of Global Epidemiology, Research Center for Zoonosis Control, Hokkaido University, N18, W9, Kita-ku, Sapporo, Hokkaido, 060-0818, Japan

    Yasuhiko Suzuki

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  1. Takashi Matsuba
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Correspondence to Takashi Matsuba.

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Matsuba, T., Suzuki, Y. & Tanaka, Y. Association of the Rv0679c protein with lipids and carbohydrates in Mycobacterium tuberculosis/Mycobacterium bovis BCG. Arch Microbiol 187, 297–311 (2007). https://doi.org/10.1007/s00203-006-0195-4

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