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Influence of Proline Substitution on the Bioactivity of Mammalian-Derived Antimicrobial Peptide NK-2

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Abstract

Multidrug-resistant bacteria are emerging as a global threat, making the search for alternative compounds urgent. Antimicrobial peptides (AMPs) became a promising hotspot due to their distinct action mechanism and possibility to be used as an alternative or complement to traditional antibiotics. However, gaining a better understanding about the relationship between antimicrobial peptides structure and its bioactivity is crucial for the development of next generation of antimicrobial agents. NK-2, derived from mammalian protein NK-lysin, has potent antitumor and bactericidal abilities. As proline was considered to be an effective α-helix breaker due to its restricted conformation, to better comprehend the effects of proline in the structure-activity relationship of NK-2, we constructed two NK-2 analogs. We examined the biological activities of NK-2 and its proline substitution analogs and analyzed the resulting conformational changes. Our results showed that introducing proline into the primary sequence of NK-2 significantly decreased the antitumor, antibacterial, and cytotoxic effects, as well as DNA binding activity by changing the α-helix content. However, α-helical content was not the only determining factor, the position of proline inserted was also critical. This study will allow for clearer insight into the role of proline in structure and bioactivity of NK-2 and provide a foundation for future studies.

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Abbreviations

AMPs:

Antimicrobial peptides

CD:

Circular dichroism.

F-moc:

N-9-fluorenylmethoxycarbonyl

RBC:

Red blood cell.

MIC:

Minimum inhibitory concentration

MTT:

3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide

TFE:

2,2,2-Trifluoroethanol

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (nos. 81601351, 81573265), the Natural Science Foundation of Gansu Province (17JR5RA267), and the research funds with the approval of the Ethical Committee of the Faculty of The First Hospital of Lanzhou University, China (no. ldyyyn2015-02).

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Author notes

  1. Jiexi Yan and Xiaolei Liang contributed equally to this work.

Authors and Affiliations

  1. The Key Laboratory, The First Hospital of Lanzhou University, Donggang Road, NO.1, Lanzhou, Gansu, 730000, People’s Republic of China

    Jiexi Yan, Xiaolei Liang, Chang Liu, Lanxia Zhou & Li Zhao

  2. Second School of Clinical Medicine, Lanzhou University, Lanzhou, 730000, China

    Yuemei Cheng

  3. School of Basic Medical Sciences, Institute of Biochemistry and Molecular Biology, Lanzhou University, Lanzhou, 730000, China

    Kairong Wang

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  1. Jiexi Yan
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Correspondence to Li Zhao.

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Yan, J., Liang, X., Liu, C. et al. Influence of Proline Substitution on the Bioactivity of Mammalian-Derived Antimicrobial Peptide NK-2. Probiotics & Antimicro. Prot. 10, 118–127 (2018). https://doi.org/10.1007/s12602-017-9335-1

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