Protein is an important concept in life, because food is full of protein, and the human body needs a lot of protein every day to maintain life. With the advancement of modern science and technology, people's research on protein is becoming more and more in-depth, and microscopic protein research results have emerged. For example, studying protein interaction is a relatively important topic. Let's take a look at what protein interaction is. Protein-protein interaction refers to the process in which two or more protein molecules form a protein complex through non-covalent bonds.Protein-protein interactions constitute a major component of the cellular biochemical reaction network and are of great significance in regulating cells and their signals. Protein interaction studies can be divided into two categories according to the experimental purpose: one is to verify whether there is an interaction between two proteins, and the other is to screen for interacting proteins of a protein of interest.
(1) The nature of the probe protein and the source of the interacting protein: The nature of the probe protein: it can be a full-length protein, an interaction-related domain, or even a smaller biologically active fragment synthesized by chemical synthesis. Sources of interacting proteins: Cell lines that are known to endogenously express the probe protein are ideal experimental materials for searching for proteins that interact with it. (2) Preparation of cell lysates: The key factors for the successful preparation of crude cell extracts include: cell lysis method; pH control; temperature; and avoiding protein degradation. These all require multiple repetitions in preliminary experiments to optimize cell lysis conditions. It is sometimes not necessary to disrupt cells to extract recombinant proteins. For example, many high-expressing mammalian cell lines (especially Chinese hamster ovary cells, CHO cells) and yeast cell lines have been engineered to secrete recombinant proteins, making it possible to purify recombinant proteins directly from cell-conditioned medium and culture filtrate. It is important to concentrate large volumes of cell-conditioned media as quickly as possible to obtain a “protease-free environment” using these methods. |
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