Recombinant Human Histidine-Rich Glycoprotein

• Cat.No.: HRG-332H
• Product Overview: Recombinant human Heregulin (HRG) protein was expressed inE. coli. MW= 44kDa (full length) and 30kDa (extracellular domain).

Specification

• Species: Human
• Source: E.coli
• Tag: Non
• Description: Heregulin (HRG) is a glycoprotein of 44kDa which binds to c-erbB-3 and c-erbB-4 receptors with low and high affinities, respectively. Heregulins constitute a family of mosaic proteins comprising of at least 12 members. They contain certain recognizable domains which include: an amino terminal (N-ter) region; an immunoglobulinlike motif (Ig); a glycosylated spacer sequence (glyco); an EGF-like motif (EGF), which is subdivided into α and β isoforms; a juxtamembrane region subdivided into isoforms 1-3; a hydrophobic transmembrane domain (TM); a common cytoplasmic tail (Cyto); and a variable-length cytoplasmic tail (a-c). Using this nomenclature, several isoforms of HRG are designated as HRG-α1, HRG-α2, HRG-α3, HRG-β1, HRG-β2, HRG-β3, HRG-α2b, etc.
• Comments: In SDS-PAGE, the purified recombinant Heregulin α1 protein appears as a single band at 30kDa. Heregulin α1 protein modulates the proliferation and differentiation of cultured mammary cells and stimulates the tyrosine kinase activity of ErbBs.
• Applications And Suggested Dilutions: Western Blotting, Stimulation of Tyrosine Kinase Activity of Erbs (Protein at 5nM); Cell Proliferation and Differentiation (Protein at 5nM: dilute 5μg of stock protein to 30ml of culture medium). The optimal dilution for a specific application should be determined by the investigator.
• Positive Control: T47-D breast cancer cells.
• Supplied As: Purified Heregulin α1 is packaged in physiological saline, pH 7.4, with 0.2% BSA. Note that this product is supplied without Sodium azide or any other preservative.
• Characterization: On SDS-PAGE commassie blue stained gel, the purified recombinant protein shows a band at 45 kDa.
• Storage And Stability: Store vial at -20°C to -70°C. When stored at the recommended temperature, this protein is stable for 24 months.
• Full Length: Full L.

Gene Information

• Gene Name: HRG histidine-rich glycoprotein [ Homo sapiens ]
• Synonyms: HRG; histidine-rich glycoprotein; HPRG; HRGP; DKFZp779H1622; thrombophilia due to elevated HRG; histidine-proline rich glycoprotein; Histidine-proline-rich glycoprotein
• Gene ID: 3273
• mRNA Refseq: NM_000412
• Protein Refseq: NP_000403
• UniProt ID: P04196
• Chromosome Location: 3q27
• MIM: 142640
• Pathway: Hemostasis
• Function: cysteine-type endopeptidase inhibitor activity; heparin binding; serine-type endopeptidase inhibitor activity

Citation

Structure and Interactions of a Dimeric Variant of sHIP, a Novel Virulence Determinant of Streptococcus pyogenes

Journal: Frontiers in Microbiology    PubMed ID: 26903974    Data: 2016/2/5

Authors: Carl Diehl, Magdalena Wisniewska, Mats Wikstr?m

Article Snippet:Subsequently, the bacteria were diluted to a concentration of 2 × 10 6 cfu/ml in 10 mM MES buffer, pH 5.5, containing 5 mM glucose.Subsequently, the bacteria were diluted to a concentration of 2 × 10 6 cfu/ml in 10 mM MES buffer, pH 5.5, containing 5 mM glucose.. Fifty microliters of the bacterial solution was incubated with recombinant His-tagged HRG (Creative BioMart) at a concentration of 0.45 μM together with various concentrations of protein sHIPwt or protein sHIPqp for 40 min at 37°C.. Serial dilutions of the incubation mixtures were plated on TH agar, plates were incubated over night at 37°C and the number of cfu's were determined.Serial dilutions of the incubation mixtures were plated on TH agar, plates were incubated over night at 37°C and the number of cfu's were determined.

Functional and Structural Properties of a Novel Protein and Virulence Factor (Protein sHIP) in Streptococcus pyogenes

Journal: The Journal of Biological Chemistry    PubMed ID: 24825900    Data: 2014/6/27

Authors: Magdalena Wisniewska, Lotta Happonen, Mats Wikstr?m

Article Snippet:Analysis of the molecular interactions between sHIP and HRG, streptococcal protein G, and human ubiquitin, respectively, was performed on a Biacore T200 instrument (GE Healthcare).Analysis of the molecular interactions between sHIP and HRG, streptococcal protein G, and human ubiquitin, respectively, was performed on a Biacore T200 instrument (GE Healthcare).. HRG (Creative BioMart), protein G (Sigma-Aldrich), or ubiquitin (Boston Biochem) was immobilized on a CM3 sensor chip using standard amine coupling at pH 5.0 in 10 m m acetate buffer at two different immobilization levels (1000 and 2000 response units).. All binding experiments were performed at 25 °C in Dulbecco's PBS buffer, pH 7.3 (Biowest) at two flow rates, 30 and 60 μl/min.All binding experiments were performed at 25 °C in Dulbecco's PBS buffer, pH 7.3 (Biowest) at two flow rates, 30 and 60 μl/min.

Biophysical studies of the interaction between sHIP and intact HRG and an HRG peptide. A, the interaction between immobilized recombinant HRG and recombinant sHIP in solution was recorded by SPR (Biacore). Experiments were carried out at 25 °C in Dulbecco's PBS buffer, pH 7.3. The normalized response units (RU) at equilibrium are plotted versus concentration of sHIP (μm). The data were fit to a 1:1 steady-state binding model, and the dissociation constant (Kd) was found to be 12 ± 1 μm. B, ITC showing the binding between protein sHIP and intact HRG at pH 5.5. C, ITC experiments showing the binding between protein sHIP and intact HRG at pH 7.5. D, ITC showing the binding between protein sHIP and HRGsp. The raw data of the experiments are presented in the top panel. The area below each injection peak is equal to the total heat released for that injection. When this integrated heat is plotted against the molar ratio of titrant added to the peptide solution in the cell, a complete binding isotherm for the interaction is obtained (bottom panel). A model for either one binding site (B) or two independent sites (C and D) was used to fit the data. The solid line is the calculated curve using the best fit parameters. The thermodynamic parameters for the interaction are shown in the bottom panel.