Abstract
In Arctic and Antarctic marine regions, where the temperature declines below the colligative freezing point of physiological fluids, efficient biological antifreeze agents are crucial for the survival of polar fish. One group of such agents is classified as antifreeze glycoproteins (AFGP) that usually consist of a varying number (n = 4–55) of [AAT] n -repeating units. The threonine side chain of each unit is glycosidically linked to β-d-galactosyl-(1 → 3)-α-N-acetyl-d-galactosamine. These biopolymers can be considered as biological antifreeze foldamers. A preparative route for stepwise synthesis of AFGP allows for efficient synthesis. The diglycosylated threonine building block was introduced into the peptide using microwave-enhanced solid phase synthesis. By this versatile solid phase approach, glycosylated peptides of varying sequences and lengths could be obtained. Conformational studies of the synthetic AFGP analogs were performed by circular dichroism experiments (CD). Furthermore, the foldamers were analysed microphysically according to their inhibiting effect on ice recrystallization and influence on the crystal habit.
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Abbreviations
- AFGP:
-
Antifreeze glycopeptide or -protein
- AFP:
-
Antifreeze protein
- Br:
-
Broad
- CD:
-
Circular dichroism
- DIPEA:
-
N,N-Diisopropylethylamine
- DMF:
-
N,N-Dimethylformamide
- DPPA:
-
Diphenylphosphoryl azide
- EtOAc:
-
Ethyl acetate
- Fmoc:
-
N-(9H-Fluoren-9-yl)-methoxycarbonyl
- Gal-GalNAc:
-
β-d-Galactosyl-(1 → 3)-α-N-acetyl-d-galactosamine
- HATU:
-
O-(7-Azabenzotriazol-1-yl)-N,N,N′N′-tetramethyluronium hexafluorophos-phate
- HOAt:
-
1-Hydroxy-7-azabenzotriazole
- MeOH:
-
Methanol
- HOBt:
-
1-Hydroxybenzotriazole
- HPLC:
-
High performance liquid chromatography
- MALDI-ToF MS:
-
Matrix assisted laser desorption ionization-time of flight mass spectrometry
- NaOMe:
-
Sodium methoxide
- NMP:
-
1-Methyl-2-pyrrolidone
- NMR:
-
Nuclear magnetic resonance
- NMM:
-
4-Methyl-2-pyrrolidinone
- PyBOB:
-
(Benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate
- SPPS:
-
Solid phase peptide synthesis
- TBDPS:
-
tert-Butyldiphenylsilyl
- TBTU:
-
O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate
- TCA:
-
Trichloroacetic acid
- TMSOTf:
-
Trimethylsilyl trifluoromethanesulfonate
- TFA:
-
Trifluoroacetic acid
- VCDC:
-
Vacuum column dry chromatography
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Acknowledgments
The authors gratefully acknowledge support from Deutsche Forschungsgemeinschaft (SFB 613, Project A8). L.N. was funded by a PhD fellowship of Bielefeld University.
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The authors declare that they have no conflict of interest.
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Nagel, L., Plattner, C., Budke, C. et al. Synthesis and characterization of natural and modified antifreeze glycopeptides: glycosylated foldamers. Amino Acids 41, 719–732 (2011). https://doi.org/10.1007/s00726-011-0937-8
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DOI: https://doi.org/10.1007/s00726-011-0937-8