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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
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
| Cat.No | ACP04852 | Target Name | LRP2 |
|---|---|---|---|
| Target Synonyms | Calcium sensor protein; DBS; Glycoprotein 330; gp330; Heymann nephritis antigen homolog; Low-density lipoprotein receptor-related protein 2; LRP-2; Lrp2; LRP2_HUMAN; Megalin | Form | Liquid or Lyophilized powder |
| Expression System | Yeast | Expression Range | 1186-1389aa |
| Mol Weight | 24.2kDa | Protein Length | Extracellular Domain |
| Purity | Greater than 90% as determined by SDS-PAGE. | Storage Buffer | 5%-50% glycerol. Lyophilized powder form: the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, Liquid form: default storage buffer is Tris/PBS-based buffer, pH 8.0. |
| Target Species | Human | Uniprot ID | P98164 |
|---|
Uniprot Id
P98164
Target Species
Human
Target Name
LRP2
Target Full Name
Low-density lipoprotein receptor-related protein 2
Target Function
Multiligand endocytic receptor. Acts together with CUBN to mediate endocytosis of high-density lipoproteins. Mediates receptor-mediated uptake of polybasic drugs such as aprotinin, aminoglycosides and polymyxin B. In the kidney, mediates the tubular uptake and clearance of leptin. Also mediates transport of leptin across the blood-brain barrier through endocytosis at the choroid plexus epithelium. Endocytosis of leptin in neuronal cells is required for hypothalamic leptin signaling and leptin-mediated regulation of feeding and body weight. Mediates endocytosis and subsequent lysosomal degradation of CST3 in kidney proximal tubule cells. Mediates renal uptake of 25-hydroxyvitamin D3 in complex with the vitamin D3 transporter GC/DBP. Mediates renal uptake of metallothionein-bound heavy metals. Together with CUBN, mediates renal reabsorption of myoglobin. Mediates renal uptake and subsequent lysosomal degradation of APOM. Plays a role in kidney selenium homeostasis by mediating renal endocytosis of selenoprotein SEPP1. Mediates renal uptake of the antiapoptotic protein BIRC5/survivin which may be important for functional integrity of the kidney. Mediates renal uptake of matrix metalloproteinase MMP2 in complex with metalloproteinase inhibitor TIMP1. Mediates endocytosis of Sonic hedgehog protein N-product (ShhN), the active product of SHH. Also mediates ShhN transcytosis. In the embryonic neuroepithelium, mediates endocytic uptake and degradation of BMP4, is required for correct SHH localization in the ventral neural tube and plays a role in patterning of the ventral telencephalon. Required at the onset of neurulation to sequester SHH on the apical surface of neuroepithelial cells of the rostral diencephalon ventral midline and to control PTCH1-dependent uptake and intracellular trafficking of SHH. During neurulation, required in neuroepithelial cells for uptake of folate bound to the folate receptor FOLR1 which is necessary for neural tube closure. In the adult brain, negatively regulates BMP signaling in the subependymal zone which enables neurogenesis to proceed. In astrocytes, mediates endocytosis of ALB which is required for the synthesis of the neurotrophic factor oleic acid. Involved in neurite branching. During optic nerve development, required for SHH-mediated migration and proliferation of oligodendrocyte precursor cells. Mediates endocytic uptake and clearance of SHH in the retinal margin which protects retinal progenitor cells from mitogenic stimuli and keeps them quiescent. Plays a role in reproductive organ development by mediating uptake in reproductive tissues of androgen and estrogen bound to the sex hormone binding protein SHBG. Mediates endocytosis of angiotensin-2. Also mediates endocytosis of angiotensis 1-7. Binds to the complex composed of beta-amyloid protein 40 and CLU/APOJ and mediates its endocytosis and lysosomal degradation. Required for embryonic heart development. Required for normal hearing, possibly through interaction with estrogen in the inner ear.
Target Involvement
Donnai-Barrow syndrome (DBS)
Target Subcellular Location
Apical cell membrane; Single-pass type I membrane protein. Endosome lumen. Membrane, coated pit. Cell projection, dendrite. Cell projection, axon.
Target Protein Families
LDLR family
Target Tissue Specificity
Expressed in first and third trimester cytotrophoblasts in the placenta (at protein level). Absorptive epithelia, including renal proximal tubules.
Target Research Area
Immunology
Target Synonyms
Calcium sensor protein; DBS; Glycoprotein 330; gp330; Heymann nephritis antigen homolog; Low-density lipoprotein receptor-related protein 2; LRP-2; Lrp2; LRP2_HUMAN; Megalin
Target Background
The protein encoded by this gene, low density lipoprotein-related protein 2 (LRP2) or megalin, is a multi-ligand endocytic receptor that is expressed in many different tissues but primarily in absorptive epithilial tissues such as the kidney. This glycoprotein has a large amino-terminal extracellular domain, a single transmembrane domain, and a short carboxy-terminal cytoplasmic tail. The extracellular ligand-binding-domains bind diverse macromolecules including albumin, apolipoproteins B and E, and lipoprotein lipase. The LRP2 protein is critical for the reuptake of numerous ligands, including lipoproteins, sterols, vitamin-binding proteins, and hormones. This protein also has a role in cell-signaling; extracellular ligands include parathyroid horomones and the morphogen sonic hedgehog while cytosolic ligands include MAP kinase scaffold proteins and JNK interacting proteins. Recycling of this membrane receptor is regulated by phosphorylation of its cytoplasmic domain. Mutations in this gene cause Donnai-Barrow syndrome (DBS) and facio-oculoacoustico-renal syndrome (FOAR).
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