The study of proteins and their function is central to understanding both cells and organisms. Proteins serve as catalysts that maintain metabolic processes in the cell, as structural elements both within and outside the cell and as signaling molecules between cells and their components.  The program consists of approximately twenty (20) laboratory sessions. This module is divided into the following sections.

 

Consistent with our aims to provide biotechnology training of international standard; the training programs begin with training in the standard requirements of working in modern biotechnology laboratories.  Students are give training in safe working practices in a biotechnology working environment.  This session is meant to help students understand how to conduct themselves in modern biotechnology laboratories and lay the foundation for subsequent lab sessions.

 

Understanding the properties of proteins and determination of protein concentrations is an essential technique in all aspects of protein and proteomics studies. Proteins differ in size, shape, isoelectric point, hydrophobicity, and biological affinity; study of these properties is necessary for understanding their structural and functional roles and their relationship in living systems.

 

This lab activity is designed to teach students the principles behind protein estimation and three of the most widely used methods in protein estimation. The first two are the alkaline copper solution methods and the third is the dye binding protein assays.

 

Students will learn about protein solubility and how it is affected by various parameters, including temperature, pH, salt and dielectric constant. They will understand about protein precipitation due to low pH, high salt and in the presence of organic solvents and about protein denaturation as a result of high temperature.

 

Students study different classes of proteins based on their solubility and function. This lab activity provides an opportunity to understand cell membrane and the characteristic of hydrophobic membrane proteins.

 

Enzymatic-proteins are functional workhorse of life and as such carry out cellular reactions in cells. Enzymes convert target molecules (substrates) into different molecules (products).

 

This lab activity involves analysis of an enzyme reaction using specific enzyme substrate and inhibitor. Students study how the rate of enzyme reaction is dependent on substrate concentration and influence of agents (inhibitors) and conditions that affects the enzyme reaction such as pH and temperature.

 

This lab activity is designed to demonstrate the dynamic nature of protein molecules in living cells, by incorporating hands-on training in protein analysis and electrophoresis. Students will perform proteolytic degradation of protein molecules and learn to understand the role and significance of proteases in normal cells and disease processes.

 

There are a number of diseases that are caused by misfolding of protein molecules such as bovine spongiform encephalopathy (BSE), Parkinson's disease and type II (non-insulin dependent) diabetes. This lab activity shows proper folding is necessary for biological activity. Researchers study an enzyme to demonstrate denaturation and renaturation of protein molecules using Zymogram.

 

Protein electrophoresis is a method of analyzing a mixture of proteins by means of gel electrophoresis. It can be modified to determine structure and function of proteins.

 

Teaches protein electrophoresis using polyacrylamide gels. Students cast polyacrylamide gels, prepare and load samples, migrate proteins by electrophoresis, and then stain the gels for visualization of the protein bands.

 

This lab activity teaches the fundamentals of protein structure. It is designed to deepen the understanding of protein molecules, using non-denaturing and denaturing electrophoresis. Students understand the difference between primary, secondary, tertiary and quaternary structures of different proteins.

 

The protein make-up of an organism is unique to that individual species, however many essential proteins are highly conserved.  This lab activity uses protein electrophoresis and protease digestion to compare and contrast the same protein from various organisms.  Students learn about conservation of genetic information in essential proteins.

 

Since proteins are the biomolecules that are important functionally and structurally, it is often valuable to be able to purify and study a protein in isolation. This ability to isolate and study a purified protein lies at the heart of much of modern biochemistry. If a protein can be isolated and purified, it can be studied in isolation from other proteins. In this module students will use different strategies employed in protein purification.

 

The lab activity exposes students to how research laboratories handle delicate samples for protein analysis and it serves as an advanced training in protein electrophoresis and protein analysis. This study involves the subcellular fractionation of a tissue sample into fractions enriched with nuclear, mitochondrial or cytoplasmic proteins.

 

This lab activity teaches common protein fractionation techniques used during protein purification, including acid and salt fractionation. This lab activity involves preparation of a crude protein extract and fractionation of proteins by incremental changes in pH and salt concentration.

 

This lab activity teaches gel filtration or size exclusion chromatography and the use of this method in purification of biological samples. This method is based on separation of protein molecules based on their molecular size. It is generally used to separate biological molecules and to determine molecular weights and molecular weight distributions of polymers.

 

This lab activity involves preparation of a crude protein extract and running ion-exchange chromatography for isolating proteins. Ion exchange chromatography is used to separate charged molecules from complex biological samples. The charged molecules bind to a solid support carrying an opposite charge to the molecule.

 

This lab activity is designed to teach students the basic principle of hydrophobic chromatography. Hydrophobic chromatography is based on the fact that protein molecules can have extensive hydrophobic regions.

 

This lab activity teaches the basic principle of affinity chromatography utilizing as highly specific affinity column. Affinity chromatography is a powerful tool for the purification of specific molecules, including proteins.