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| What is Proteomics? |
Proteomics is a study of complete protein present in any living organism. Proteomics provides analysis of the structure, composition, function, and expression of proteins within the organism. Applications of proteomics have significantly impacted drug discovery, the environment, and the agricultural sector. Proteomics has made a revolution in the field of science and medicine. Proteomics is basically of three types structure, function, and expression.
What is Proteomics?
Proteomics is a large-scale study of proteins including their structure, composition, and function within the biological system. Proteomics plays a role in understanding the complexities of cellular functions, signaling pathways, and disease mechanisms within the living organism. Proteomics is used to reveal:
· Protein relation and interaction within the genome of a person
· How Proteins function in metabolic and biological processes
T Types of Proteomics
There are three basic types of proteomics
Expression Proteomics
Expression protein is a technique in proteomics to assess
both qualitative and quantitative expression of proteins. It is used to
evaluate protein expression between affected and non-affected patients. Expression
proteomics is used to detect the expression of proteins in different tissues and
cells.
Structural Proteomics
As the name indicates it is used to identify the structure
of proteins and its complexities within the protein. It includes the whole three-dimensional
structure of proteins such as membrane, nucleus, and every cellular organelle.
Functional Proteomics
Functional Proteomics determines the molecular pathway and
protein function within the cell. This type of proteomics defines protein
interaction by identifying unknown proteins from a protein complex involved in
molecular mechanisms.
Process of Proteomics
The first and foremost step is the isolation of protein from the sample. Cells or tissues are isolated and proteins are removed by preserving
their native state. Next, protein
separation is achieved through gel electrophoresis and liquid chromatography.
These techniques separate proteins based on their size, charge, and affinity. Basically, there are two methods of proteomics:
Top-Down Proteomics
In top-down proteomics, whole protein is subjected to mass
spectrometry analysis without treating proteins by enzymes. This method preserves the native structure of the protein and allows direct analysis of post-translational modifications. The
process involves sample preparation, Mass spectrometry, and data
interpretation.
Bottom-Up Proteomics
In a bottom-up approach, enzymes first digested proteins into peptides and then subjected to mass spectrometry analysis. This
method is adopted widely due to its compatibility with high-throughput
workflows and its ability to identify a large number of peptides. The method
involves protein digestion, protein separation, and mass spectrometry.
Proteomics: Techniques
Gel Electrophoresis 2D
2D gel electrophoresis is a technique that separates large
complexes of protein based on their charge-to-mass ratio. Proteins are
separated based on two properties: In the first dimension by charge using an isoelectric point and in the second dimension by size using SDS-PAGE. This
technique helps in identifying variations in protein expression patterns. 2D gel electrophoresis uses a fluorescent dye to
differentiate more than two protein samples on the same gel.
Antigen-antibody methods
Techniques known as ELISA, western blotting identifies
specific protein complex and quantify their expression level. In Elisa, a specific
antigen binds to the particular antibody attached to a protein on the surface. Western
blotting uses gel electrophoresis to separate proteins and then transfer them to the membrane where it is treated with primary antibody and secondary antibody-producing enzyme-substrate complex. This technique is also used for
quantitative analysis of protein.
Mass Spectrometry
Mass Spectrometry is a quantitative technique that
determines mass to charge ratio of ions. In proteomics, mass spectrometry
identifies and quantifies proteins by measuring masses of peptides generated
from protein digestion. Proteins first
get ionized by an ion source in a mass spectrometer, and then these ions are
separated by a mass analyzer and detected.
Liquid Chromatography
Liquid chromatography coupled with mass spectrometry enables the separation and analysis of complex protein mixtures. Liquid chromatography breaks down peptides into proteins based on properties like hydrophobicity, or
charge, which is then subjected to a mass spectrometer for identification.
Protein Microarray
This is a high-throughput technique that consists of a solid
surface or membrane where numerous arrays of proteins are immobilized. A signal is produced upon the reaction of the fluorescently labeled probe with immobilized
protein. Protein Microarray enables protein-protein interactions,
antigen-antibody interaction, and enzymatic reactions.
Applications of Proteomics
Biomarker Identification
One of the most prominent applications of proteomics lies in
discovering biomarkers. Researchers can detect particular proteins
associated with certain diseases by comparing proteome profiles of both healthy
and diseased individuals. In cancer research, proteomics has enabled in
identification of biomarkers that help in providing early detection, diagnosis, and
monitoring treatment response.
Personalized Medicine
Proteomics has played a crucial role in providing personalized treatment to individual patients based on their molecular profiles. By analyzing the proteomic profile of a
patient, clinicians can determine treatment strategies and predict how a
patient might respond to a particular therapy. This approach has become successful
in selecting targeted remedies for diseases like cancer depending on their molecular
features.
Drug Development and Target Identification
Drug discovery involves targeting specific proteins involved
in disease mechanisms. Proteomics enables the identification of potential drug targets and helps in assessing the effect of
drugs on protein expression and interactions. Researchers can analyze how drugs
influence protein, along with their mechanism and
potential side effects through techniques such as mass spectrometry.
Environmental and Agricultural Applications
Besides the medical field, proteomics has also entered agriculture and environmental science. In agriculture, proteomics enhances crop yields and quality by detecting proteins associated with traits like disease resistance, nutritional content, and other environmental-resistant features. In environmental studies, proteomics plays a role in monitoring the impact of pollutants on organisms and ecosystems. It also provides an understanding of ecological stressors and potential biomarkers of environmental toxicity.
Functional Proteomics
Protein function plays a vital role in interpreting complex
biological processes. Functional proteomics involves studying roles, protein interaction, and their operations within the cellular network. Techniques
such as protein-protein interaction, mapping, post-translational modification,
and subcellular localization studies depict how proteins contribute to cellular
processes.
Conclusion
Proteomics has developed our understanding of proteins and
their role in various biological contexts, leading to numerous applications that
extend from disease diagnosis, and treatment to drug development and
environmental monitoring. Proteomics has contributed to new discoveries and
advancements in science and medicine leading to breakthroughs in personalized medicine and precision therapeutics.
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