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Protein synthesis by solid-phase chemical ligation using a safety catch linker

Academic Article
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Overview

authors

  • Brik, A.
  • Keinan, Ehud
  • Dawson, Philip

publication date

  • June 2000

journal

  • Journal of Organic Chemistry  Journal

abstract

  • The native chemical ligation reaction has been used extensively for the synthesis of the large polypeptides that correspond to folded proteins and domains. The efficiency of the synthesis of the target protein is highly dependent on the number of peptide segments in the synthesis. Assembly of proteins from multiple components requires repeated purification and lyophilization steps that give rise to considerable handling losses. In principle, performing the ligation reactions on a solid support would eliminate these inefficient steps and increase the yield of the protein assembly. A new strategy is described for the assembly of large polypeptides on a solid support that utilizes a highly stable safety catch acid-labile linker. This amide generating linker is compatible with a wide range of N-terminal protecting groups and ligation chemistries. The utility of the methodology is demonstrated by a three-segment synthesis of vMIP I, a chemokine that contains all 20 natural amino acids and has two disulfide bonds. The crude polypeptide product was recovered quantitatively from the solid support and purified in 20%-recovered yield. This strategy should facilitate the synthesis of large polypeptides and should find useful applications in the assembly of protein libraries.

subject areas

  • Amino Acid Sequence
  • Chemistry, Organic
  • Disulfides
  • Indicators and Reagents
  • Macrophage Inflammatory Proteins
  • Molecular Sequence Data
  • Peptide Fragments
  • Proteins
  • Sepharose
  • Viral Proteins
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Identity

International Standard Serial Number (ISSN)

  • 0022-3263

Digital Object Identifier (DOI)

  • 10.1021/jo000346s

PubMed ID

  • 10864771
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Additional Document Info

start page

  • 3829

end page

  • 3835

volume

  • 65

issue

  • 12

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