The Protein Man's Blog | A Discussion of Protein Research

Detecting Properties of Protein Using Protein Binding Assay (Part 1 of 3)

Posted by The Protein Man on Sep 3, 2013 5:00:00 AM
The Protein Man

Question:

How to use Protein Binding Assay to Detect Properties of Proteins?

The Protein Man Says:

protein binding assayProteins play a major role in most life processes. They are responsible for a great number of biological processes which include cell growth and proliferation, gene expression, morphology, motility, nutrient uptake and apoptosis, to name a few. However, since the basic characteristics of proteins are highly complex, understanding their functions in the proper biological context can be quite a challenge. To do this,we may need to shift our focus from studying single proteins to investigating their interactions with other proteins. And this is where protein binding assay comes in quite handy.

The Importance of Studying Protein-Protein Interactions

Aside from studying individual protein sequence and structures, expression profile, post-translational modifications and intracellular localization, you also need to investigate and analyze their interactions with other proteins. By knowing the functions of their binding partners, you will definitely get to know your proteins of interest a lot better. In fact, this is how a lot of previously unknown proteins have been discovered.

Studying protein-protein interactions is very important since it can:

  • Help you understand a protein's function and behavior,
  • Predict biological processes that may result from the interaction of two or more proteins, and
  • Help you understand biological processes and pathways a lot better.

Types of Protein-Protein Interactions and Their Biological Effects

Protein-protein interactions can be classified as either stable or transient. While stable interactions form stable complexes, transient interactions are more temporary in nature. However, these transient interactions are the ones that control most cellular processes.

Once a set of conditions are met, binding protein partners may interact with each other and bring about a wide range of cellular processes which may include cell cycling, protein modification, folding, transport and signaling.Proteins may bind with other proteins at specific binding domains through electrostatic interactions or salt bridges, hydrogen bonding, hydrophobic bonding and dipole-dipole interactions or van der Waals forces.

There are a number of biological effects that may result from the interaction of two or more proteins. Some interactions may alter the kinetic properties of an enzyme, create a new binding site or result in the production of an intended end product while some may serve a regulatory function in an upstream or downstream event. There are also some that may change the properties of a protein, inactivate it or totally destroy it.

Now that you understand why determining protein-protein interactions is extremely important, we will be discussing the many ways by which these interactions can be analyzed by using the appropriate protein binding assay in our next post.

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Topics: Protein Concentration

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Ellyn Daugherty's Biotechnology: Science for the New Millennium

CB™ PROTEIN ASSAY: A Bradford Protein Assay

CB Protein Assay Graph

An improved Coomassie Dye based protein assay based on the Bradford Protein Assay. This assay is suitable for the simple and rapid estimation of protein concentration. This assay is based on a single Coomassie dye based reagent. The binding of protein to the dye results in a change of color from brown to blue. The change in color density is proportional to protein concentration. Protein estimation can be performed using as little as 0.5µg protein.

Features

  • Sensitivity: Linear responses over the range of 0.5µg-50µg protein
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