2. Contents
• Proteomics.
• Role of Proteomics.
• Branches of Proteomics.
• Techniques of Proteomics.
• Complexities of Proteomics.
• Advantages of Proteomics.
• Biomarkers.
4. Definition:
That’s just not a protein
biochemistry !
Proteome :
It is the complement protein found in
a single cell in a particular environment./ is
complete collection of proteins encoded by
genome of an organism.
Proteomics :
It is the study of composition,
structure, function and interaction of the proteins
directing the activities of each living cell.
5. • To study the structure and
function of protein.
• To study the 3D structure of
protein.
• Study of qualitative and
quantitative analysis of proteins.
Role of Proteomics
6.
7. Helps to identify newly discovered genes
and drug interaction
Structural proteomics
12. The first one is called
isoelectric focusing
(IEF) which separates
proteins according to
isoelectric points (pI) The second step is SDS-
polyacrylamide gel
electrophoresis
(SDSPAGE) which
separates proteins based
on the molecular weights
•Thus, thousands of
proteins can be
separated, and the
information about IEF
and molecular weights
can be obtained
Steps
13. •proteins are amphoteric
molecules and the positive,
negative, or zero net charge
they carry depending on the
pH of the surroundings.
•The isoelectric point (pI) is
defined as the pH of a solution
at which the net charge of the
protein becomes zero.
Isoelectric Focusing (IEF)
14. • A protein with a positive net charge will migrate toward
the cathode, becoming less positively charged until
reaching its pI. While a protein with a negative net
charge will migrate toward the anode, becoming less
negatively charged until it also reaches its pI.
15. Isoelectric Focusing (IEF)
• A protein mixture is
loaded at the basic end of
the pH gradient gel.
• After applying an electric
field, the proteins are
separated depending on
charges, focusing at
positions where the pl
value is equivalent to the
surrounding pH.
• Larger proteins will move
more slowly through the
gel, but with sufficient
time will catch up with
small proteins of equal
charge
16. • Be performed on flatbed or vertical systems on a
slab gel.
• The second dimension is often performed by
SDS-PAGE), which is an electrophoretic method.
• This technique is used to separate proteins by
their molecular weight.
• SDS negatively charged detergent used to
denature proteins.
SDS-PAGE
(SDS-polyacrylamide gel electrophoresis)
17. How does an SDS-PAGE Seperation Work?
• Negatively charged proteins move to positive Electrode.
• SDS coated larger proteins migrate slowly through the Gel
Matrix.
• SDS coated smaller proteins
migrate quickely through the Gel Matrix.
18.
19. Results
Silver staining:
• Sensitive and non-radioactive method.
• Silver staining is suitable for low protein
levels because of its sensitivity.
20. Coomassie Blue staining
• A relatively simple method and more
quantitative than silver staining.
• It is suitable to detect protein bands
containing about 0.2 μg or more proteins.
21. MASS SPECTROMETRY
• While 2D- gel electrophoresis
separates proteins, it doesn’t identify
them.
• MS is used to identify them which
separates charged particles or ions
according to mass.
22. 3 Major Parts
• Sours ionized the sample.
• Analyzer separate the ions on m/z ratio.
• Detector sees the ions and analyzed the
result.
23. How does a Mass Spectrometer
work?
• Samples easier to manipulate if ionised.
• Separation in analyser according to mass-to-
charge ratios (m/z).
• Detection of separated ions and their relative
abundance.
• Signals sent to data system and formatted in
a m/z spectrum .
24.
25. •The study of proteins is very
complex because the
concentration of protein is
different in each organism and in
each cell of the organism.
Complexities in proteomics
26.
27. • Shows that genetic alterations are not
the reason for all types of diseases.
• Helps in determining the proper
treatment of diseases.
• With the help of three dimensional
analysis of proteins we have found that
HIV protease is the enzyme which is
responsible for AIDS.
Advantages of study of proteomics
28. • Advanced screening for disease.
• One of the most important use of
proteomics in diagnosis is the
identification of biomarkers.
• The study of drugs in proteomics
is called pharmacoproteomics
29. Biomarkers
• Biomarkers are molecules that
indicate normal or abnormal
process taking place in your body
and may be a sign of an underlying
condition or disease.
• DNA (genes), proteins or hormones,
can serve as biomarkers, since they
all indicate something about your
health.
