General Techniques for Biomolecular Characterization

Abstract

Nucleic acids and proteins are biomolecules which are made up of chains of building blocks called nucleotides and amino acids, respectively. The synthesis of proteins is itself catalyzed by proteins and enzymes, however this process is directed by genetic material in the cell, DNA. Two types of nucleic acids occur in cells: deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Both these biomolecules, nucleic acids and proteins play very important and versatile roles in living systems. DNA acts as a carrier of genetic information whereas proteins which are translated from mRNA perform various functions in the biological processes. They are the important constituent of cell membrane, function as enzymes, antibodies and form different tissues in the body. Primary structure of a biomolecule is the linear arrangement in which monomers are joined together. Secondary and tertiary structure of these biomolecules are stabilized by a number of factors like hydrogen bonding, stacking interactions, ionic interactions, vander waal interactions etc. A detailed information about the formation of these structures, DNA/protein-ligand interaction, type of changes occurring during the enzymatic reactions can be studied by various biochemical and biophysical techniques. This review covers the basic principle of techniques like Gel-Electrophoresis, UV-Thermal Denaturation and Circular dichroism (CD) employed for the characterization of nucleic acids and proteins. Gel-Electrophoresis is an analytical tool, used by biochemists for the separation, purification and for information on structural status of biomolecules. UV-Thermal Denaturation and Circular dichroism (CD) are useful in structural studies of nucleic acids, proteins and nucleoproteins. UV-thermal denaturation is used for determining the stability of biomolecules. CD spectra of nucleic acids and proteins arise primarily from the spatial symmetry of the nucleotides and amino acids respectively, in the backbones of the macromolecules.