Journal article

A simple approach for the preparation of mature human relaxin-3

Xiao Luo, Ya-Li Liu, Sharon Layfield, Xiao-Xia Shao, Ross AD Bathgate, John D Wade, Zhan-Yun Guo

PEPTIDES | ELSEVIER SCIENCE INC | Published : 2010

Abstract

Relaxin-3 (also known as INSL7) is the most recently identified member of the insulin-like family. It is predominantly expressed in the nucleus incertus of the brain and involved in the control of stress response, food intake, and reproduction. In the present work, we have established a simple approach for the preparation of the mature human relaxin-3 peptide. We first designed and recombinantly expressed a single-chain relaxin-3 precursor in E. coli cells. After purification by immobilized metal ion affinity chromatography, refolding in vitro through disulfide reshuffling, and digestion by endoproteinase Asp-N, mature human relaxin-3 was obtained in high yield and at low cost. Peptide mappi..

View full abstract

Grants

Awarded by National Natural Science Foundation of China


Awarded by National Basic Research Program of China (973 Program)


Awarded by NHMRC


Funding Acknowledgements

This work was supported by the National Natural Science Foundation of China (30970609) and the National Basic Research Program of China (973 Program no 2010CB912604) The studies at the HFI (Melbourne) were supported in part by NHMRC project grants (#508995 and 509048) to JDW and RADB We thank Linda Chan (FNI) for the amino acid analysis and Tania Ferraro for the activity assays