This document discusses microbial proteomics and various proteomics techniques. It describes structural proteomics which analyzes protein structures to identify gene functions and interaction sites. Interaction proteomics analyzes protein-protein interactions to determine functions. Expression proteomics identifies proteins differentially expressed between related samples like healthy and diseased tissue. Microbial proteomics studies microorganism proteins using techniques like mass spectrometry, electrospray ionization, and matrix-assisted laser desorption-ionization. Gel-based separation techniques including 1D, 2D, and 3D gel electrophoresis are also discussed.

1. MICROBIAL PROTEOMICS
BY:-
T.ARUNA
(U14BR001)
DEPT OF GENETIC ENGINEERIG

2. PROTEOMICS:
 Proteomics is the large-scale study of
proteins, particularly their structures and
functions. Proteins are vital parts of living
organisms, as they are the main
components of the physiological
metabolic pathways of cells.

3. PROTEOMICS:

4. STRUCTURAL PROTEOMICS
 Structural proteomics includes the analysis of
protein structures at large-scale. It compares
protein structures and helps identify functions
of newly discovered genes.
 The structural analysis also helps to
understand that where drugs bind to proteins
and also show where proteins interact with
each other.

5. INTRACTION PROTEOMICS:
 Interaction proteomics is the analysis of protein
interactions at larger scale. The characterization
of protein-protein interactions are useful to
determine the protein functions and it also
explains the way proteins assemble in bigger
complexes.
 Technologies such as affinity purification, mass
spectrometry, and the yeast two-hybrid system
are particularly useful in interaction proteomics.

6. EXPRESSION PROTEOMICS:
 Expression proteomics includes the analysis
of protein expression at larger scale. It helps
identify main proteins in a particular sample,
and those proteins differentially expressed in
related samples—such as diseased vs.
healthy tissue.
 If a protein is found only in a diseased
sample then it can be a useful drug target or
diagnostic marker.

7. MICROBIAL PROTEOMICS:
 In order for microbiologists to embrace the
potential that proteomics offers, an
understanding of a variety of analytical tools
is required.
 MICROBIAL PROTEOMICS is a study which
deals with the analysis of proteins present in
the micro organisms using various methods.

8. MASS SPECTROMETRY:
 It is nothing but the usage of mass
spectrometers over other analytical instruments
is that it affords a high degree of accuracy and
sensitivity ,when determining the molecular
weight of biological compounds .
 A mass spectrometer is an instrument that
produces ions from a sample, separates them
according to their mass-to-charge ratio (m/z)
and records the relative abundance of each of
the ions to obtain a mass spectrum .

9. MASS SPECTROMETRY:

10. ELECTROSPRAY IONISATION (ESI)
 Electrospray ionization (ESI) is a technique used
in mass spectrometry to produce ions using
an electrospray in which a high voltage is applied to a
liquid to create an aerosol. It is especially useful in
producing ions from macromolecules because it
overcomes the propensity of these molecules to
fragment when ionized.
 ESI is different than other atmospheric pressure
ionization processes (e.g. MALDI) since it may
produce multiply charged ions, effectively extending
the mass range of the analyser to accommodate
the kDa-MDa orders of magnitude observed in
proteins and their associated polypeptide fragments

11. ELECTROSPRAY IONISATION:

12. MATRIX-ASSISTED LASER DESORPTION-IONISATION
(MALDI)
 Matrix-assisted laser
desorption/ionization (MALDI) is a
soft ionization technique used in mass spectrometry,
allowing the analysis
of biomolecules (biopolymers such
as DNA, proteins, peptidesand sugars) and
large organic molecules , which tend to be fragile and
fragment when ionized by more conventional
ionization methods.
 It is similar in character to electrospray
ionization (ESI) in that both techniques are relatively
soft ways of obtaining ions of large molecules in the
gas phase, though MALDI produces far fewer multiply
charged ions.

13. MATRIX-ASSISTED LASER DESORPTION-
IONISATION (MALDI):

14. GEL-BASED SEPARATION

15. 1D GEL ELECTROPHORESIS:
 One-dimensional gel electrophoresis (1-DE) is
used in resolving relatively simple protein
mixtures, usually obtained after purification of
the desired protein fraction.
 In 1-DE the proteins are separated according to
their molecular weight (MW). 2-DE has become
a standard separation method in gel-based
proteomics, enabling the simultaneous
separation and visualization of thousands of
proteins

16. 1D GEL ELECTROPHROSIS:

17. 2D GEL ELECTROPHORESIS:
 2D gel electrophoresis (2DE) is a key
technique for purifying individual proteins
from complex samples based on their
isoelectric points and molecular weights.
 Simple enough in theory, but as the plethora
of protocols and articles shows, 2DE
demands patience and meticulous
optimization.

18. 2D GEL ELECTROPHORESIS:

19. 3D GEL ELECTROPHROSIS:
 3D-gel electrophoresis is a new approach to DNA and
protein analysis according to
one, two, or three independent separation
parameters
 The 3D-gel may also be used for high-throughput
analysis with two separation parameters,
e.g., comparative analysis of protein aggregates by
blue native electrophoresis and
subsequent SDS-PAGE .
 Several neurodegenerative diseases
involve amyloid-like protein aggregation.
 Our technique could provide a diagnostic picture
during their development and allows the screening of
drug candidates for their effects on
protein aggregation.

20. 3D GEL ELECTROPHROSIS: