PROTEOMICS, BIOMOLECULAR IDENTIFICATION AND GENE

THERAPY

Prepared by:

Waqar , Sarmad, Amir, Rizwan & Junaid

PHARMACISTS OF

SHAH ABDUL LATIF UNIVERSITY KHAIRPUR SINDH,PAKISTAN

INTRODUCTION TO PROTEOMICS

Proteomics is simply defined as the Study of Proteome. The term proteome was created by

Wasinger et al. (1995) and Wilkins et al. (1996) and defined as the cell- and tissue-specific patterns of

proteins expressed by the genome of an organism. Proteomics is also defined as the study of total protein content of a cell or that of an organism.

SCOPE OF PROTEOMICS

Proteomics helps in understanding the alteration in

protein expression during different stages of life cycle or

under stress condition.

Proteomics is also important with regards to drug

development against several diseases and

understanding various biological processes, as proteins

are the most favourable targets for various drugs.

SCOPE OF PROTEOMICS

Proteomics helps in understanding the structure and

function of different proteins as well as protein-protein

interactions of an organism and a minor defect in either

protein structure, its function or alternation in

expression pattern can be easily detected using

proteomics studies.

Protein to gene can be predicted by Proteomics.

CLASSIFICATION OF PROTEOMICS

1. Functional Proteomics: Identifications of Protein-Protein,

Protein-DNA and Protein-RNA interactions affecting

function.

2. Structural Proteomics: Identification of all interactions by

metal ions, toxins, drugs etc. affecting Protein structure.

3. Differential Proteomics: Determination of differences in

Protein expression.

BIO-MOLECULAR IDENTIFICATION

Initial step in all proteomic studies is the separation of

proteins mixture by Two Dimensional Gel Electrophoresis

technique, based on their individual charges in 1D. The gel is then

turned 90 degrees to separate proteins based on the difference in

their size. This separation occurs in 2nd dimension hence the

name 2D. The spots obtained in 2D electrophoresis are excised

and further subjected to mass spectrometric analysis of each

protein present in the mixture.

STEPS IN PROTEOMIC ANALYSIS

Purification of proteins: It involves extraction of protein

sample followed by purification using density gradient

centrifugation, chromatographic techniques (affinity etc.)

Separation of proteins: 2D gel electrophoresis is applied for

separation of proteins on the basis of their isoelectric points in

one dimension and molecular weight on the other. Spots are

detected using fluorescent dyes or radioactive probes.

STEPS IN PROTEOMIC ANALYSIS

Identification of proteins: The separated protein spots

on gel are excised and digested in gel by a protease

(e.g. trypsin). The eluted peptides are identified using mass

spectrometry.

Validation of Proteins: Determined amino acid sequence

is finally compared with available database to validate the

proteins.

OVERVIEW OF STEPS

TECHNIQUES INVOLVED IN PROTEIN IDENTIFICATION

GENE THERAPY

Pathological changes within an organism arise from

protein alterations. Proteomics provides a powerful set of tools

for the large-scale study of protein expression. The identification

of such changes by proteomics is essential to define drug

targets, therapeutic proteins, and disease biomarkers. Gene

therapy, primarily based on the delivery of a “therapeutic” gene

to the diseased organ, is an approach whose effect is finally due

to the function of the therapeutic protein encoded by the

GENE THERAPY

“therapeutic” gene, proteomics provides the basis required for

the design and application of gene therapies. Because in certain

diseases the absence of certain cellular or biological factors, for

example Enzymes occur and it is may be due to a gene being

defective. Cancer, infectious diseases, cardiovascular diseases,

neurological disorders and some inherited conditions are among

the areas into which the Gene therapy research is being carried

out.

APPLICATIONS OF PROTEOMICS

1.For discovery of protein biomarkers.

2.In the Study of Tumor Metastasis.

3.In the Field of Neuro-trauma.

4.In disease diagnosis.

5.In Neurology.

6.In Treatment of Autoimmune disease.

7.In Nutrition Research.