[PDF][PDF] Convergent chemical synthesis of histone H2B protein for the site-specific ubiquitination at Lys34.
P Siman, SV Karthikeyan, M Nikolov… - Angewandte Chemie …, 2013 - pure.mpg.de
Angewandte Chemie-International Edition, 2013•pure.mpg.de
Synthesis of H2B (97-125): The synthesis was carried out using Fmoc-SPPS on Rink amide
resin (0.2 mmol/g, 0.1 mmol scale). The peptide synthesis was performed on peptide
synthesizer in presence of 4 equiv of amino acid (AA), 8 equiv of DIEA and 4 equiv of
HBTU/HOBt to the initial loading of the resin. Cleavage from the resin: A mixture of TFA,
triisopropylsilane (TIS) and water (95: 2.5: 2.5) was added to the dried peptide-resin and the
reaction mixture was shaken for 2 h at RT. The resin was removed by filtration and was …
resin (0.2 mmol/g, 0.1 mmol scale). The peptide synthesis was performed on peptide
synthesizer in presence of 4 equiv of amino acid (AA), 8 equiv of DIEA and 4 equiv of
HBTU/HOBt to the initial loading of the resin. Cleavage from the resin: A mixture of TFA,
triisopropylsilane (TIS) and water (95: 2.5: 2.5) was added to the dried peptide-resin and the
reaction mixture was shaken for 2 h at RT. The resin was removed by filtration and was …
Synthesis of H2B (97-125): The synthesis was carried out using Fmoc-SPPS on Rink amide resin (0.2 mmol/g, 0.1 mmol scale). The peptide synthesis was performed on peptide synthesizer in presence of 4 equiv of amino acid (AA), 8 equiv of DIEA and 4 equiv of HBTU/HOBt to the initial loading of the resin.
Cleavage from the resin: A mixture of TFA, triisopropylsilane (TIS) and water (95: 2.5: 2.5) was added to the dried peptide-resin and the reaction mixture was shaken for 2 h at RT. The resin was removed by filtration and was washed with TFA (2 x 2 mL). To precipitate the peptide, the combined filtrate was added drop-wise to 10-fold volume of cold ether followed by centrifugation, decanting of ether and by dissolution of residue in acetonitrile-water for the HPLC purification step.
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