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Antimicrobial lipopeptaibol trichogin GA IV: role of the three Aib residues on conformation and bioactivity

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Abstract

The lipopeptaibol trichogin GA IV is a natural, non-ribosomally synthesized, antimicrobial peptide remarkably resistant to the action of hydrolytic enzymes. This feature may be connected to the multiple presence in its sequence of the non-coded residue α-aminoisobutyric acid (Aib), which is known to be responsible for the adoption of particularly stable helical structures already at the level of short peptides. To investigate the role of Aib residues on the 3D-structure and bioactivity of trichogin GA IV, we synthesized and fully characterized four analogs where one or two Aib residues are replaced by l-Leu. Our extensive conformational studies (including an X-ray diffraction analysis) and biological assays performed on these analogs showed that the Aib to l-Leu replacements do not affect the resistance to proteolysis, but modulate the bioactivity of trichogin GA IV in a 3D-structure related manner.

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Abbreviations

Aib:

α-Aminoisobutyric acid

AMPs:

Antimicrobial peptides

CF:

5(6)-Carboxyfluorescein

CFU:

Colony forming unit

Ch:

Cholesterol

DIEA:

N,N′diisopropylethylamine

DMEM:

Dulbecco’s minimal Eagle medium

DMSO:

Dimethylsulfoxide

DOPC:

1,2-dioleyl-sn-glycero-3-phosphocholine

EDC:

N-ethyl, N′-[3-(dimethylamino)propyl]carbodiimide

ESI:

Electrospray ionization

Fmoc:

Fluorenyl-9-methyloxycarbonyl

HaCaT:

Human adult low-calcium high temperature

HATU:

O-(7-azabenzotriazol-1-yl)-1,1,3,3 tetramethyluronium hexafluorophosphate

HBTU:

O-(benzotriazol-1-yl)-1,1,3,3 tetramethyluronium hexafluorophosphate

HFIP:

1,1,1,3,3,3-hexafluoroisopropanol

HMBC:

Heteronuclear multiple bond coherence

HMQC:

Heteronuclear multiple quantum coherence

HOAt:

7-aza-1-hydroxy-benzotriazole

HOBt:

1-hydroxy-benzotriazole

hRBCs:

Human red blood cells

Lol:

Leucinol

MeOH:

Methanol

MES:

2-(N-morpholino) ethanesulfonic acid

MIC:

Minimal inhibitory concentration

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium

nOct:

n-Octanoyl

PBS:

Phosphate buffered saline

RMSD:

Root-mean-square deviation

SDS:

Sodium dodecylsulphate

SPPS:

Solid-phase peptide synthesis

SUVs:

Small unilamellar vesicles

TFA:

Trifluoroacetic acid

Z:

Benzyloxycarbonyl

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Acknowledgments

The authors are grateful to Prof. A. Dolmella (Department of Pharmaceutical Sciences, University of Padova) for granting access to the Gemini diffractometer and his help with X-ray diffraction data collection and processing. Financial support for the acquisition of the Agilent Technologies Gemini diffractometer was provided by the University of Padova through the 2008 “Scientific Equipment for Research” initiative.

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Authors and Affiliations

  1. ICB, Padova Unit, CNR, Department of Chemistry, University of Padova, 35131, Padova, Italy

    Marta De Zotti, Barbara Biondi, Marco Crisma, Claudio Toniolo & Fernando Formaggio

  2. Department of Biotechnology, Chosun University, 501-759, Gwangju, Korea

    Yoonkyung Park

  3. Department of Cellular Molecular Medicine, School of Medicine, Chosun University, 501-759, Gwangju, Korea

    Kyung-Soo Hahm

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  1. Marta De Zotti
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  2. Barbara Biondi
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  3. Yoonkyung Park
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  4. Kyung-Soo Hahm
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  5. Marco Crisma
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  6. Claudio Toniolo
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  7. Fernando Formaggio
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Correspondence to Marta De Zotti.

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De Zotti, M., Biondi, B., Park, Y. et al. Antimicrobial lipopeptaibol trichogin GA IV: role of the three Aib residues on conformation and bioactivity. Amino Acids 43, 1761–1777 (2012). https://doi.org/10.1007/s00726-012-1261-7

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