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 language | English |
|---|---|
| Pages (from-to) | 124-130 |
| Number of pages | 7 |
| Journal | FEBS Letters |
| Volume | 402 |
| Issue number | 2-3 |
| DOIs | |
| State | Published - Jan 27 1997 |
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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 journal › Article
}
TY - JOUR
T1 - Expression, purification and characterization of recombinant human proinsulin
AU - Cowley, Darrin J.
AU - Mackin, Robert
PY - 1997/1/27
Y1 - 1997/1/27
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0031052016&partnerID=8YFLogxK
U2 - 10.1016/S0014-5793(96)01511-6
DO - 10.1016/S0014-5793(96)01511-6
M3 - Article
C2 - 9037180
AN - SCOPUS:0031052016
VL - 402
SP - 124
EP - 130
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 2-3
ER -