Analysis Biomolecular Interactions of TC and PC with ErbB1 on Epithelial Cells by BLI (CAT#: STEM-MB-0243-CJ)

Introduction

The epidermal growth factor receptor (EGFR, or ErbB-1) subfamily is comprised of four transmembrane receptor tyrosine kinases, ErbB-1, -2, -3, and -4. These receptors have been implicated in the pathogenesis of various human cancers. The significance of ErbB-2 and/or ErbB-1 overexpression has been studied extensively in breast and lung cancer. Overexpression of ErbB1 in conjunction with autocrine ligand production has also been associated with decreased survival. Differences in the total amount and ratio of glycoprotein complexes have been detected in various HCMV strains, potentially influencing the variability in cellular tropism.

Structural and biochemical characterization of HCMV TC and PC by mass spectrometry and mutagenesis analysis revealed that TC and PC form two mutually exclusive cell entry complexes. These complexes are formed by disulphide bonds between the cysteine 144 of gL and either the cysteine 351 of gO or the cysteine 162 of pUL128, meaning that both complexes use the same binding domain at the N terminus of gH/gL.

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Principle Applications Procedure Materials Notes Related Products

Principle

Bio-Layer Interferometry (BLI) is an optical technique for measuring macromolecular interactions by analyzing interference patterns of white light reflected from the surface of a biosensor tip. BLI experiments are used to determine the kinetics and affinity of molecular interactions. In a BLI experiment, one molecule is immobilized to a Dip and Read Biosensor and binding to a second molecule is measured. A change in the number of molecules bound to the end of the biosensor tip causes a shift in the interference pattern that is measured in real-time.

Applications

Immunology/Inflammation; Virology

Procedure

1. Detect Buffers and prepare samples. BLI experiments are set up with one molecule immobilised on the surface of the biosensor (load sample) and a second molecule in solution (the analytical sample).
2. Fix the load sample on the biocompatible biosensor while the analytical sample is in solution.
3. The biosensor tip is immersed in the solution so that the target molecule begins to bind to the analysis sample.
4. Set up and run the BLI experiment. Molecules bound to or dissociated from the biosensor can generate response curves on the BLI system; unbound molecules, changes in the refractive index of the surrounding medium or changes in flow rate do not affect the interferogram pattern.
5. Collect and analyse data on the BLI's system.

Materials

• Equipment: Fortebio Bio-Layer Interferometry (BLI)
• Sample Type: DNA, RNA, Protein, Antibodies, Peptides, Small Molecules
• Optionals: Epithelial cells, Minimum essential medium Eagle

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