Expression, purification and characterization of recombinant human proinsulin

Darrin J. Cowley, Robert Mackin

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

We have recently developed a method to produce native human proinsulin using a bacterial expression system. A proinsulin fusion protein was recovered from inclusion bodies and cleaved using cyanogen bromide. The released proinsulin polypeptide was S-sulfonated and purified by anion exchange chromatography. Following refolding, proinsulin was purified by reversed-phase high-performance liquid chromatography. Combined peptide mapping and mass spectrometric analysis indicated that the proinsulin contained the correct disulfide bridging pattern. This proinsulin will be used to study the specificity of the furin/PC family of converting enzymes by using it as a substrate in a recently developed assay.

Original languageEnglish
Pages (from-to)124-130
Number of pages7
JournalFEBS Letters
Volume402
Issue number2-3
DOIs
StatePublished - Jan 27 1997

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Proinsulin
Purification
Furin
Cyanogen Bromide
Peptides
Peptide Mapping
Inclusion Bodies
High performance liquid chromatography
Reverse-Phase Chromatography
Chromatography
Population Groups
Disulfides
Anions
Assays
Fusion reactions
High Pressure Liquid Chromatography
Substrates
Enzymes
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Expression, purification and characterization of recombinant human proinsulin. / Cowley, Darrin J.; Mackin, Robert.

In: FEBS Letters, Vol. 402, No. 2-3, 27.01.1997, p. 124-130.

Research output: Contribution to journalArticle

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