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Expression, Purification, and Therapeutic Implications of Recombinant sFRP1 by Archita Ghoshal; Siddhartha Sankar Ghosh is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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Secreted frizzled-related proteins (sFRPs) constitute a family of proteins, which impede the Wnt signaling pathway. Upregulation of the Wnt cascade is one of the multiple facets of carcinogenesis. Herein, we report the expression, solubilization, purification, characterization, and anti-cell proliferative activity of a novel recombinant GST-tagged sFRP1 of human origin. sFRP1 was cloned into pGEX-4T2 bacterial expression vector, and the recombinant protein was overexpressed in Escherichia coli BL21 (DE3). It was solubilized from inclusion bodies with N-lauroylsarcosine and Triton X-100, before being purified to homogeneity using glutathione agarose affinity chromatography column. The purified protein was characterized using Western blotting, MALDI TOF-TOF, and circular dichroism spectroscopy analysis. Homology modeling and docking studies revealed that tagging GST with sFRP1 does not change the binding conformation of the cysteine-rich domain and hence, possibly does not alter its function. The novel anti-proliferative activity of GST-sFRP1 was demonstrated in a dose-dependent manner on two cancer cell lines, viz., HeLa (cervical cancer) and MCF-7 (breast cancer). Also, combination

Who reads Expression, Purification, and Therapeutic Implications of Recombinant sFRP1?

Author

Archita Ghoshal; Siddhartha Sankar Ghosh

Publisher

Springer Science and Business Media LLC

Published

2014

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)