Microbial proteomics helps to identify the proteins associated with microbial activity, microbial host-pathogen interactions, and antimicrobial resistant mechanism. Microbial activity of pathogens can be confirmed by using the 2-D gel-based and gel-free method with the combination of MALDI-TOF-LC-MS/MS.

1. Presentation topic: Microbial
Proteomics.

2. What is Proteomics?
 A ”proteome” is a complete set of proteins produced in an
organism, system, or biological context during their
lifetime.
 “Proteomics” is the large-scale study of proteomes i;e to
study the dynamic protein products of the genome and their
interaction, and large scale characterization of the proteins
expressed by a genome.
 Why proteomics?
 The behavior of gene and gene product is difficult or
impossible to predict from gene sequence.
 Even if gene is transcribed, its expression maybe regulated
at the level of translation.

3. Proteomics: The new era of Microbiology.
 It is now more than 10 years since the publication of the first
microbial genome sequence and science is now moving towards
a post genomic era with transcriptomics and proteomics
offering insights into cellular processes and function.
 Rapid advancement in current technologies such as mass
spectrometry, bioinformatics and protein separation
technologies have produced a step change in our current
proteomic capabilities
and
number of groups are taking advantage of this cutting edge
technology to discover more about the physiology and
metabolism of microorganisms.

4. Progress of Proteomics in Microbiology.

5. Techniques used in Microbial proteome
analysis.
 In the last decades, characterization and differentiation of
microbial proteome was performed by means of gel-based
and gel-free protein separation methods.
 Numerous techniques for the analysis of proteome exist
and they can be divided in two major groups;
1- Spectrophotometric methods, which comprise
HPLC (High-performance liquid chromatography) and
LC/MS (Liquid chromatography-mass spectrometry);
2- Antibodies-based methods, which include ELISA
(Enzyme-linked immunosorbent assay),
immunoprecipitation, immunoelectrophoresis and western
blot.

6. Specification of Techniques.

7. Classical proteomics workflow to study physiology
of microbes.

8. Mass Spectrometry based bacterial
proteomics.

9. Gel-Based Proteomics.

10. Applications Of Proteomics

11.  Quantification/Identification of proteins is done
through various proteomics techniques.
 Body fluid is passed over a chip, or subject to
other chromatography-based capture device.
 Retained peptides or proteins are identified by
either single or tandem mass spectrometry.
 MS pattern differences of diseased from healthy
individual samples serve as biomarker for that
disease.
 Hence, further used for the prognosis of disease.
Biomarker Discovery by Proteomics:

12. Drug Target Identification by
Proteomics.
 Drug, or a library of candidates drugs, is tethered
to a solid support or chip.
 Extract from cell, tissues or fluid is then washed
over the chip solid support and interaction
proteins adhere to the drug bait(Capture molecule
for drug).
 The complex can be washed and adhering
proteins can be eluted.
 The eluate undergo proteomics analysis and all
the binding partners are identified.

13. Target Validation by Proteomics:
 Examine protein changes as a consequence of
a specific drug treatment.
 Focus on expressions differences, signaling
cascade,transcriotional or secretory event for
specific proteins involved in drug treatment.
 Evaluate specific protein- protein interactions
and effect of particular drug on this
interaction.
 Examine the consequences of a particular
drug treatment on representative proteins that
have been selectively isolated.

14. Future Development
Ensuring the characterization of proteome of Complex
Organisms and we are a long off potentially 90% away
from the entire proteome of Complex living organism.
However important results would be achieved soon by
utilizing high-throughput techniques.
This is an emerging field and that
the technology only starting to
move from the development phase
into real applications.
So far, we are just capable of characterizing the tip of the
proteo-berg. To better grasp the proteome complexity, we need
better analytical as well as chemometric and statistical tools
that are capable of refining and rationalizing the large amount
of data that we are collecting.

15. Thank You!