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Chemical Synthesis of 19F-labeled HIV-1 Protease using Fmoc-Chemistry and ChemMatrix Resin

  • Bruce Merrifield Commemorative Issue
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International Journal of Peptide Research and Therapeutics Aims and scope Submit manuscript

HIV-1 protease (PR) has been extensively studied due to its importance as a target in AIDS therapy. The enzyme can be obtained via expression of its cloned gene in an appropriate system, or via chemical synthesis. We required a reliable source of fluorine-labeled HIV-1 protease for NMR studies. As our attempts to incorporate a labeling step in overexpression experiments in E. coli failed, we turned to chemical synthesis. Herein is described the first chemical synthesis of an active, 99 amino acid residue HIV-1 encoded protease using Fmoc-chemistry on a total PEG-based resin (CM resin), and labeled with 19F at the Phe residue. Also reported here are NMR studies of the labeled synthetic protein with a synthetic dimerization inhibitor.

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Abbreviations

CM:

ChemMatrix resin

DCM:

dichloromethane

DMF:

N,N-dimethylformamide

DIEA:

N,N-diisopropylethylamine

DTT:

dithiothreitol

FDA:

Food and drug administration

Fmoc:

9-fluorenylmethoxycarbonyl

HATU:

1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo-[4,5-b]pyridinium hexafluorophospate 3-oxide

HIV-1 PR:

HIV-1 protease

HOAt:

1-hydroxy-7-azabenzotriazole (3-hydroxy-3H-1,2,3-triazolo-[4,5-b] pyridine

MALDI:

matrix-assisted laser desorption ionization

MeCN:

acetonitrile

MSNT:

(1-(mesitylene)-2-sulfonyl-3-nitro-1H-1,2,4-triazole)

MTBE:

Methyl tert-butyl ether

NMI:

1-(methylimidazole)

NMR:

nuclear magnetic resonance

PEG:

poly(ethyleneglycol)

RP-HPLC:

reversed-phase high performance liquid chromatography

RT:

reverse transcriptase

TBTU:

1-[bis(dimethylamino)methylene]-1H-benzotriazolium tetrafluoroborate 3-oxide

TFA:

trifluoroacetic acid

TOF:

time-of-flight

UV:

ultraviolet

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Acknowledgments

This work was supported by MCYT-FEDER (Bio2005-00295 and GEN2003-20642-C09-04), and Generalitat de Catalunya (CERBA and 2005SGR-00663).

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Authors and Affiliations

  1. Institut de Recerca Biomèdica, Parc Científic de Barcelona, C/ Josep Samitier, 1-5, Barcelona, E-08028, Spain

    Silvia Frutos, Judit Tulla-Puche, Fernando Albericio & Ernest Giralt

  2. Departament de Química Orgànica, Universitat de Barcelona, Martí i Franquès, 1, Barcelona, E-08028, Spain

    Fernando Albericio & Ernest Giralt

Authors
  1. Silvia Frutos
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  2. Judit Tulla-Puche
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  3. Fernando Albericio
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  4. Ernest Giralt
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Corresponding author

Correspondence to Ernest Giralt.

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Frutos, S., Tulla-Puche, J., Albericio, F. et al. Chemical Synthesis of 19F-labeled HIV-1 Protease using Fmoc-Chemistry and ChemMatrix Resin. Int J Pept Res Ther 13, 221–227 (2007). https://doi.org/10.1007/s10989-006-9070-z

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  • DOI: https://doi.org/10.1007/s10989-006-9070-z

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