A. The overall objective of this project is to discover and develop novel or potentially important bioactive proteins of significant biological and medical importance from sustainable resources in Malaysia. Thus, the two main areas that have been identified to initiate the Malaysian proteomic works are:
i. The identification and exploitation of bioactive proteins of clinical importance
ii. The identification and exploitation of novel bioactive marine and animal toxins1. The identification and exploitation of bioactive proteins of clinical importance
Serum proteins are routinely use as markers for clinical diagnosis. The up- and/or down-regulation of proteins expressed in the serum like the acute-phase proteins, immunoglobulins immune modulators, cell growth and differentiation factors and regulators, non immune cell defense factors and polypeptide hormones facilitates diagnosis of various clinical disorders. The advent of proteomic technology provides a novel method to evaluate the use of other serum proteins as diagnostic or prognostic markers as well as a tool to evaluate progression of a disease and/or the effectiveness of a treatment. The presence of unique proteins in the sera of patients of a certain disease or clinical manifestation can be easily detected by separating the serum samples using 2-dimensional gel electrophoresis and comparing its resolved pattern with that obtained from normal healthy control. These proteins can subsequently identified by using the MALDI-TOF PRO mass spectrometry. Unlike other existing methodology, the proteomics technology is capable of performing the analysis of all serum proteins simultaneously. The technology has been applied in this project to screen the sera of patients for possible the possible presence of novel human serum proteins of diagnostic/prognostic importance. The diseases that have been analysed are:
i. breast carcinoma
ii. cervical carcinoma
iii. ovarian carcinoma
iv. nasopharyngeal carcinoma
v. congenital hypothyroidism
vi. diabetes mellitus type I2. The identification and exploitation of novel bioactive marine and animal toxins
There is also a tremendous effort in harvesting protein data from natural proteins obtained from various sources using the methodologies of proteomics. Among these natural bioreactive proteins of interest are the various types of toxins such as cardiotoxins and neurotoxins obtained from animal, plant and microbials. In addition, most of the toxins are accompanied by a variety of bioreactive factors such as anti-coagulant, anti-thrombotic, and anti-platelet binding factors together with active enzymes including phospholipases, endo- and exo-nucleotidases, hyaluronidase, neuraminidase, protease and many others. As Malaysia is rich in its natural resources, the availability of many venomous and poisonous fauna and flora provide a pool of starting material for study.2.1. Among the organisms that will be included in this part of the study are snakes venoms, jellyfish venoms, poisonous fishes/bivalves Saxitoxins. The proteins profile of the secretion and venom produced by these organisms will be studied on the 2-D electrophoresis and subsequent identification of proteins using mass spectrometry (MALDI-TOF PRO). The identification and characterization of the protein patterns can also be used as a protein-map of the organisms for species identification and will also be useful in the elucidation of the binding sites of action. Further kinetics studies and thus be carried out to determine the affinity and potency of these bioreactive proteins at their targets. Useful antibodies can be developed for neutralization of envenomation cases.
2.2. A variety of frog and toad species have been identified to secrete compounds that have potential clinical uses, such as treatment of chronic and acute pain, control of narcotic addiction and alcohol abuse, and the suppression of the immune response during transplant surgery.
EXECUTIVE SUMMARY
The
Application of Proteomics Technology in the Identification and
Exploitation of Novel Bioactive Proteins.
Since no investigation on these bioactive peptides from frog dermal secretions have been conducted thus far in Malaysia, this part of the project involves with identifying frog species in Malaysia for potential chemical prospecting by screening all compounds found in each secretion. The environmental influence of the skin secretion of the same species from multiple localities in certain selected states will also be investigated.
2.3. The classifications of microbial strains are based upon the analysis of chemical constituents either as unique biomarkers or as contributors to distinct ratios of chemical components. In this context, the hospital-acquired infections at the KL and Penang General hospitals have been a prevalent problem. In this initiative, microorganisms will be isolated at these sites and subjected to mass spectral analysis. Revelation of biomarkers molecules including lipids, phospholipids, lipopolysaccharides, oligosaccharides, proteins and DNA will be used as the basis for the development of chemotaxonomy of the organisms as well as identification of potential drug targets. Outer membrane proteins from the bacterial cell wall component will also be studied as they have been shown to trigger immune response in the host. This eventually can lead to the elucidation of new drug targets not only to direct towards the antigenic epitope regions but also in managing resistant-strains-bacteria. Drugs or peptide drugs will be tested in animal serum to investigate its suitability as a vaccine in such infections.
B. Participating Institutions:-
i. Professor Iekhsan Othman; iekhsan@um.edu.my, University Malaya
ii. Associate Professor Onn Haji Hashim; onnhashim@um.edu.my; University of Malaya
iii. Encik Ismail Ishak; anasofiah@hotmail.com; Fisheries Research Institute, Penang
iv. Puan Faazaz Abdul Latif; faazaz@tm.net.my; Fisheries Research Institute, Subang
v. Dr. Shahrul Anuar Mohd Sah; sanuar@usm.my; Malaysian Science University, Penang
vi. Professor Aishah Abdul Latif; aishah@dcc.usm.my; Malaysian Science University, Penang
Period of the project : 3 Years (2001 - 2004)Achievements
Progress/Achievements for year one
Discovery
1. Serum proteome maps of patients with breast cancer, cervical carcinoma and congenital hypothyroidism were generated. Based on the maps, some bioactive proteins from the patients were visually and quantitatively different form their respective normal controls. Specific proteins that are elevated or inhibited in the expression process were determined by comparing with Swiss Expasy Map and would further be confirmed using Maldi-Tof Pro and protein sequencing.
2. The proteome maps of patients with coronary heart disease and diabetes mellitus were initiated.
3. The 2-D electrophoretic protein profiles of secretions and venoms were also being analyzed. The protein maps of distinctive species obtained are used as a species identification.
- 7 Publication
Status
- On-going