Decidual prolactin-related protein: heterologous expression and characterization

Endocrinology
C A RasmussenM J Soares

Abstract

As a first step in understanding the role of decidual PRL-related protein (dPRP) during pregnancy, we have generated recombinant dPRP protein. In this report, we present data on the generation, purification, and characterization of recombinant dPRP protein. The dPRP complementary DNA was subcloned into the pMSXND vector, and the vector was transfected into Chinese hamster ovary (CHO) cells by electroporation. After appropriate selection, amplification, and induction procedures, recombinant dPRP was purified from conditioned medium of the CHO-dPRP cells using ultrafiltration, size-exclusion chromatography, and reverse phase HPLC. Recombinant dPRP was found to possess electrophoretic mobility, immunoreactivity, and N-terminal amino acid sequence identical to those of dPRP isolated from decidual tissue. Polyclonal antibodies were generated to the recombinant dPRP and used for Western blot analysis. dPRP is capable of binding heparin, and a significant fraction of synthesized dPRP resides within the decidual extracellular matrix. Recombinant dPRP failed to bind to PRL receptors and showed no stimulatory activity in the PRL-dependent rat Nb2 lymphoma cell proliferation assay. Additional studies have shown that heterologous expressio...Continue Reading

Citations

May 10, 2008·The Journal of Biological Chemistry·S M Khorshed AlamMichael J Soares
Jul 9, 2004·Reproductive Biology and Endocrinology : RB&E·Michael J Soares
Mar 16, 2007·Biochemical Genetics·Ying LiYa-Ping Zhang
Jun 20, 1998·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·H MüllerM J Soares
Jan 18, 2006·Microscopy Research and Technique·Karin CarboneTelma M T Zorn
May 23, 2001·Biology of Reproduction·H C Keith HoB P Schick
Dec 19, 2002·Endocrinology·Dustin O WiemersMichael J Soares
Nov 7, 2017·Birth Defects Research·Michael J SoaresKeisuke Kozai
May 20, 2008·Archives of Physiology and Biochemistry·Yang YuReinald Fundele
Dec 23, 1999·The Journal of Biological Chemistry·A Prigent-TessierG Gibori
Dec 21, 2006·Differentiation; Research in Biological Diversity·Umashankar SinghReinald H Fundele

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