Synthetic, structural and biological studies of the ubiquitin system: The total chemical synthesis of ubiquitin

R. Ramage, J. Green, T. W. Muir, O. M. Ogunjobi, S. Love, K. Shaw

Research output: Contribution to journalArticlepeer-review

105 Scopus citations

Abstract

The small protein ubiquitin (76 amino acids) has been synthesized under optimized conditions by Merrifield solid-phase methodology using the N(α)-Fmoc protecting group. The crude polypeptide mixture was purified to homogeneity by gel filtration, dialysis and a combination of cation- and anion-exchange chromatography to yield ubiquitin. Amino acid analysis, enzymic digestion and sequencing by automated Edman degradation were used to authenticate the primary structure. Isoelectric focusing and m.s. were used to demonstrate that the final product was greater than 98% pure with a final yield of 93 mg (4.3%) from a single synthesis on a 0.25 nmol scale.

Original languageEnglish (US)
Pages (from-to)151-158
Number of pages8
JournalBiochemical Journal
Volume299
Issue number1
DOIs
StatePublished - 1994
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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