Biomolecular Structure ID: Polarimetry and Circular Dichroism
Optical Rotatory Dispersion (ORD) and Circular Dichroism (CD) are two important techniques widely used in the study of biomolecules, particularly the structure of proteins and nucleic acids. These techniques are based on the interaction of molecules with light, specifically how the molecules affect plane-polarized light that passes through them.
Optical Rotatory Dispersion
1. Fundamentals
ORD is based on the optical activity of chiral molecules towards plane-polarized light. When plane-polarized light passes through a solution containing chiral molecules, the plane of polarization rotates. This rotation, known as optical rotation, is measured by a polarimeter, typically in degrees.
2. Applications
ORD is commonly used to determine the absolute and relative configurations of compounds, monitor changes in chiral centers during chemical or biological processes (such as enzymatic reactions), and assess the purity and concentration of chiral drugs.
3. Information Content
ORD primarily provides information about chiral centers, such as their absolute configuration and chemical environment, but does not offer detailed information about the precise structure of the molecule.
Circular Dichroism (CD)
1. Fundamentals
CD is based on the differential absorption of left- and right-circularly polarized light by chiral molecules. The difference in absorption of these two types of light gives rise to a CD signal, which can be used to analyze the structure and conformation of molecules.
2. Applications
CD is widely used in the structural study of proteins and nucleic acids, determining the secondary structure of these macromolecules, studying protein folding, understanding molecular interactions (like protein-ligand or protein-protein interactions), and monitoring how environmental changes (such as pH, solvent, etc.) affect the structure of molecules.
3. Information Content
A CD spectrum can provide detailed information about the secondary structure of the molecule, such as the relative proportions of α-helices, β-pleated sheets, and random coils, and can also be used to study the kinetics and interactions of molecules.
Applications
1. Protein Secondary Structure Analysis
CD spectroscopy can be used to determine the different types of secondary structure present in a protein, and monitor changes in these structures under various environmental conditions (such as different pH, temperature, or presence/absence of cofactors). This is crucial for understanding the function and stability of proteins.
2. Protein Folding and Conformational Changes
CD spectroscopy can be used to study intermediate states in the protein folding process, as well as structural changes in proteins upon interacting with ligands or other proteins. This information is extremely important for drug development and understanding the biological functions of proteins.
3. Nucleic Acid Structure Studies
CD is also used for studying the structure of nucleic acids, especially for distinguishing between different forms of DNA (such as A-, B-, and Z-DNA).
4. Enantiomeric Purity and Chiral Drug Analysis
Optical activity analysis is the standard method in the pharmaceutical industry for determining the enantiomeric purity of drugs. Since different enantiomers can have very different pharmacological activities or metabolic properties, this analysis is critical for the safety and effectiveness of drugs.
Advantages and Disadvantages of the Techniques
1. Advantages
(1) Non-Destructiveness
The sample can be used for further studies after measurement.
(2) Sensitive to Conditions
They can monitor the effect of pH, temperature, ionic strength, etc. on molecular structure.
(3) Speed and Sensitivity
Especially CD spectroscopy, which can quickly provide information about the secondary structure of molecules.
2. Disadvantages
(1) Limited Information
They typically cannot provide detailed information about the tertiary or quaternary structure of molecules.
(2) Requiring Prior Knowledge
Understanding and interpreting CD spectra require some prior knowledge about the system being studied.
(3) Requirements for Sample Purity and Concentration
Impurities and insufficient sample concentration may affect the accuracy of results.
With further research and technological advancements, optical activity and CD spectroscopy are becoming more powerful and diverse tools. Future developments may include combining them with other types of spectroscopy and structural biology techniques to provide more detailed information about complex biomolecular systems. Simultaneously, with the advancement of machine learning and data analysis techniques, these methods are expected to become more precise and intelligent, playing larger roles in areas such as biomolecular research, drug design, and disease diagnosis.
How to order?