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Studies on the assembly of a leucine zipper antibacterial peptide and its analogs onto mammalian cells and bacteria

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Abstract

Membrane-interaction and assembly of a leucine zipper peptide (LZP), and its single (SASA) and double (DASA) alanine-substituted analog onto mammalian, hRBCs and 3T3 cells and bacteria, Escherichia coli and Staphylococcus aureus were studied as a model system to understand the plausible role of assembly on their contrasting cytotoxic but similar bactericidal activities. Peptides’ ability to depolarize and damage the membrane organization of hRBC and 3T3 cells decreased from LZP to SASA and to DASA which may be related to their decrease in assembly onto these mammalian live cells and oligomerization states in the presence of these cell membranes or zwitterionic PC/Chol lipid vesicles. However, LZP and its analogs showed appreciable similarities in damaging or depolarizing the E. coli or S. aureus cells, which further matched with their comparable assembly and oligomerization either onto these live cells or the cell membranes or in the presence of negatively charged PC/PG lipid vesicles.

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Abbreviations

hRBCs:

Human red blood cells

Fmoc:

N-(9-Fluorenyl) methoxycarbonyl

Chol:

Cholesterol

HPLC:

High performance liquid chromatography

CFU:

Colony forming unit

PC:

Egg phosphatidylcholine

PG:

Egg phosphatidylglycerol

NBD:

4-Fluoro-7-nitrobenz-2-oxa-1,3-diazole

MALDI-TOF:

Matrix-assisted laser desorption ionization time-of-flight

PBS:

Phosphate-buffered saline

MICs:

Minimum inhibitory concentrations

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

FRET:

Fluorescence resonance energy transfer

LUV:

Large unilamellar vesicles

FITC:

Fluorescein isothiocyanate

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Acknowledgments

The work was supported by the CSIR network project NWP 0005. The authors are extremely thankful to A. L. Vishwakarma for recoding the flow cytometry profiles. Brijesh Kumar Pandey is acknowledged for his contribution in preparation of revised version of the manuscript. A. A and S.A. acknowledge the receipt of a senior and junior research fellowship from CSIR, India, respectively.

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Authors and Affiliations

  1. Molecular and Structural Biology Division, Central Drug Research Institute, CSIR, Lucknow, 226001, India

    Aqeel Ahmad, Sarfuddin Azmi & Jimut Kanti Ghosh

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  1. Aqeel Ahmad
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  2. Sarfuddin Azmi
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  3. Jimut Kanti Ghosh
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Correspondence to Jimut Kanti Ghosh.

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The CDRI communication number of this manuscript is 7202.

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Ahmad, A., Azmi, S. & Ghosh, J.K. Studies on the assembly of a leucine zipper antibacterial peptide and its analogs onto mammalian cells and bacteria. Amino Acids 40, 749–759 (2011). https://doi.org/10.1007/s00726-010-0744-7

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