Journal article

Design and recombinant expression of insulin-like peptide 5 precursors and the preparation of mature human INSL5

Xiao Luo, Ross AD Bathgate, Wei-Jie Zhang, Ya-Li Liu, Xiao-Xia Shao, John D Wade, Zhan-Yun Guo

AMINO ACIDS | SPRINGER | Published : 2010

Abstract

Insulin-like peptide 5 (INSL5) is a recently identified insulin superfamily member. Although it binds to and activates the G-protein coupled receptor, RXFP4, its precise biological function remains unknown. To help determine its function, significant quantities of INSL5 are required. In the present work, three single-chain INSL5 precursors were designed, two of which were successfully expressed in E. coli cells. The expressed precursors were solubilized from inclusion bodies, purified almost to homogeneity by immobilized metal-ion affinity chromatography, and then refolded in vitro. One precursor could be converted to two-chain human INSL5 bearing an extended N-terminus of the A-chain (desig..

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Grants

Awarded by National Basic Research Program of China


Awarded by National Natural Science Foundation of China


Awarded by NHMRC


Funding Acknowledgements

This work was supported by the National Basic Research Program of China (973 Program, no. 2010CB912604) and by the National Natural Science Foundation of China (30970609, 30700124). 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 analyses and Sharon Layfield (FNI) for the bioassays.