Expression, purification and preliminary crystallographic analysis of human thyroid hormone responsive protein

Abstract

Thyroid hormone responsive protein (Thrsp, also known as Spot 14 and S14) is a carbohydrate-inducible and thyroid-hormone-inducible nuclear protein specific to liver, adipose and lactating mammary tissues. Thrsp functions to activate genes encoding fatty-acid synthesis enzymes. Recent studies have shown that in some cancers human Thrsp (hS14) localizes to the nucleus and is amplified, suggesting that it plays a role in the regulation of lipogenic enzymes during tumourigenesis. Thrsp, a member of the Spot 14 superfamily, is an acidic homodimeric protein with no sequence similarity to other mammalian gene products and its biochemical function is elusive. To shed light on the structure-function relationship of this protein, human Thrsp was crystallized. Recombinant human Thrsp (hThrsp), the N-terminally truncated human Thrsp 10-146 (hThrsp9) and their selenomethionyl (SeMet) derivatives were expressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. Diffraction-quality crystals were grown at 293 K using Li 2 SO 4 as a precipitant. Using synchrotron radiation, data for the hThrsp SeMet derivative, hThrsp9 and its SeMet derivative were collected to 4.0, 3.0 and 3.6 Å resolution, respectively, at 100 K. The crystals of full-length hThrsp and its SeMet derivative belonged to space group P4 1 2 1 2, with approximate unit-cell parameters a = b = 123.9, c = 242.1 Å , = = = 90.0 . In contrast, the crystals of the truncated hThrsp9 and its SeMet derivative belonged to space group P2 1 2 1 2 1 , with approximate unit-cell parameters a = 91.6, b = 100.8, c = 193.7 Å , = = = 90.0 . A molecular-replacement solution calculated using a murine Spot 14 structure as a search model indicated the presence of six molecules per asymmetric unit, comprising three hThrsp homodimers.

Weighted gene correlation network analysis identifies the critical long non-coding RNAs participate in the progression of osteosarcoma

“…PLIN1 was also reported to of prognostic significance in human breast cancer and might be a potentially new gene therapy target (Zhou et al 2016). THRSP is a thyroid-hormone-inducible and carbohydrate-inducible and protein, functions to activate genes involved in fatty-acid synthesis enzymes (Zhang et al 2011). Evidence suggested it plays a role in tumorigenesis by regulating lipogenic enzymes (Kinlaw et al 2006).…”