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Characterization of the Interactions of Lysozyme with DNA by Surface Plasmon Resonance and Circular Dichroism Spectroscopy

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Abstract

Association with nucleic acid has been recognized as a unique role of lysozyme and may explain why lysozyme was called a killer protein against HIV infection. In the present study, we characterized the interactions of lysozyme and its derived peptides with a biotin-labeled pUC19 plasmid DNA. Real-time detection of the macromolecular interaction was performed using the SPR (surface plasmon resonance) spectroscopy. The SPR sensorgrams were analyzed and the association and dissociation rate constants as well as the dissociation equilibrium constant K D were, thus, estimated. The results reveal that other than the electrostatic interactions between the basic protein and the nucleotide sequences carrying negative charges, the specific DNA-binding motifs at the N- and C-termini of lysozyme were also involved in the interactions. The nonapeptide RAWVAWRNR (aa 107–115 of lysozyme) reported previously to block HIV-1 viral entrance and replication was also able to bind DNA with its K D value comparable to that of histones. The possibilities of ligand-binding-induced conformational changes were investigated using the circular dichroism spectroscopy. The CD spectra (200–320 nm) reveal that the conformational changes indeed occur as the spectra of lysozyme–DNA interactions are much less at the major trough region than the sum of individual spectra. The interaction of lysozyme with DNA molecules may interfere with DNA replication, modulate gene expression, and block bacterial and viral infections. These all suggest that human lysozyme may represent part of the innate immune system with a very broad protective spectrum.

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Abbreviations

CD spectroscopy:

circular dichroism spectroscopy

HEW:

hen egg-white

SPR:

surface plasmon resonance

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Acknowledgement

The present work was supported in part by the grants (NSC95-2113-M001-043 to LSK; NSC94-2311-B259-005 to DS) from National Science Council, ROC. The authors also thank Academia Sinica for providing the research supporting grant (Academia Sinica 72v112) for visiting scholars (to DS).

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Authors and Affiliations

  1. Department of Life Science, Institute of Biotechnology and the Bioinformatic Program, National Dong Hwa University, Hualien, Taiwan, 974, Republic of China

    Kuo-Chih Lin & David Shiuan

  2. College of Life Science, National Tsinghua University, Hsinchu, 300, Taiwan

    Ming-Tsai Wey

  3. Institute of Chemistry, Academia Sinica, Taipei, Taiwan, 112, Republic of China

    Ming-Tsai Wey & Lou-Sing Kan

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  1. Kuo-Chih Lin
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  2. Ming-Tsai Wey
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  3. Lou-Sing Kan
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  4. David Shiuan
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Correspondence to David Shiuan.

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Lin, KC., Wey, MT., Kan, LS. et al. Characterization of the Interactions of Lysozyme with DNA by Surface Plasmon Resonance and Circular Dichroism Spectroscopy. Appl Biochem Biotechnol 158, 631–641 (2009). https://doi.org/10.1007/s12010-008-8348-3

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  • DOI: https://doi.org/10.1007/s12010-008-8348-3

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