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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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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DOI: https://doi.org/10.1007/s00726-012-1261-7