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Synthesis and characterization of natural and modified antifreeze glycopeptides: glycosylated foldamers

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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.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Organic and Bioorganic Chemistry, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany

    Lilly Nagel, Carolin Plattner & Norbert Sewald

  2. Physical Chemistry, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany

    Carsten Budke, Thomas Berkemeier & Thomas Koop

  3. Institute of Chemistry, Eötvös Loránd University, P.O. Box 32, 1518, Budapest 112, Hungary

    Zsuzsanna Majer

  4. Department of Animal Biology, University of Illinois at Urbana-Champaign, 524 Burrill Hall, 407 S Goodwin, Urbana, IL, 61801, USA

    Arthur L. DeVries

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  1. Lilly Nagel
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  2. Carolin Plattner
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  3. Carsten Budke
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  4. Zsuzsanna Majer
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  5. Arthur L. DeVries
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  7. Thomas Koop
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  8. Norbert Sewald
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Correspondence to Norbert Sewald.

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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

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