To produce recombinant human low-density lipoprotein receptor-related protein 2 (LRP2) in yeast, the gene encoding the extracellular domain of the human LRP2 protein (1186-1389aa) is co-inserted into an expression vector with an
N-terminal 6xHis-tag gene and transformed into yeast cells. The yeast cells are grown under conditions that promote protein expression. After sufficient growth, the cells are lysed to release the recombinant LRP2 protein. The recombinant human LRP2 protein is purified from the cell lysate through affinity chromatography. The purity of the protein is assessed using SDS-PAGE, up to 90%.
The Human LRP2 protein, also known as megalin, is a large transmembrane glycoprotein receptor that plays a crucial role in various physiological processes [1]. It functions as a multifunctional cell surface receptor involved in mediating the uptake of hormones and vitamins bound to carrier proteins, such as vitamin D-binding protein, retinol-binding protein, and sex hormone-binding globulins [2]. LRP2 acts as an auxiliary receptor for sonic hedgehog (SHH) signaling, controlling the internalization and cellular trafficking of SHH complexes [3].
Studies have highlighted the significance of LRP2 in different developmental processes, including cardiac development and neural tube closure. Loss of LRP2 has been associated with heart defects in mice, emphasizing its crucial role in cardiac development [4]. Furthermore, LRP2 has been implicated in neural tube closure, with its deficiency leading to neural tube defects in mouse embryos, a condition that can be prevented by folic acid supplementation [5].
References:
[1] , Megalin-mediated trafficking of mitochondrial intracrines: relevance to signaling and metabolism,, vol. 3, no. 6, 2021. https://doi.org/10.33696/immunology.3.118
[2] E. Kur, N. Mecklenburg, R. Cabrera, T. Willnow, & A. Hammes, Lrp2 mediates folate uptake in the developing neural tube, Journal of Cell Science, 2014. https://doi.org/10.1242/jcs.140145
[3] A. Christ, A. Christa, E. Kur, O. Lioubinski, S. Bachmann, T. Willnowet al., Lrp2 is an auxiliary shh receptor required to condition the forebrain ventral midline for inductive signals, Developmental Cell, vol. 22, no. 2, p. 268-278, 2012. https://doi.org/10.1016/j.devcel.2011.11.023
[4] M. Baardman, M. Zwier, L. Wisse, A. Groot, W. Kerstjens-Frederikse, R. Hofstraet al., Common arterial trunk and in lrp2 knock out mice indicate a crucial role of lrp2 in cardiac development, Disease Models & Mechanisms, 2016. https://doi.org/10.1242/dmm.022053
[5] J. Sabatino, B. Stokes, & I. Zohn, Prevention of neural tube defects in lrp2 mutant mouse embryos by folic acid supplementation, Birth Defects Research, vol. 109, no. 1, p. 16-26, 2017. https://doi.org/10.1002/bdra.23589
