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Genomic translations of Fenugreek ( Trigonella foenum-graecum ) to derive its proteomic insights

Genomic translations of Fenugreek ( Trigonella foenum-graecum ) to derive its proteomic insights. Geetika Jethra, Priya Gupta, Jyoti Mihra, Alka Pawar and Sharda Choudhary. Presented By : Geetika Jethra Young Profession II National Research Centre on Seed Spices Tabiji , Ajmer. Seed Spices.

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Genomic translations of Fenugreek ( Trigonella foenum-graecum ) to derive its proteomic insights

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  1. Genomic translations of Fenugreek (Trigonellafoenum-graecum) to derive its proteomic insights Geetika Jethra, Priya Gupta, Jyoti Mihra, Alka Pawar and Sharda Choudhary Presented By: GeetikaJethra Young Profession II National Research Centre on Seed Spices Tabiji, Ajmer

  2. Seed Spices • Indian spices include a variety of spices grown across the Indian subcontinent and mostly categorised under Apicaeae family. • Spices are used for flavour, colour, aroma and preservation of food or beverages in almost all of the Indian Cuisines. • But the database available on Apicaeae and Fabaceaefamily is sparsely populated with sequences.

  3. Statistics of Seed Spices

  4. Fenugreek (Trigonellafoenum-graecum) • Fenugreek is commonly known as methiin Hindi • It is an important leguminous spices and well known aromatic and medicinal herb. • Fenugreek is used as both seed and leaf. • Fenugreek seed contains carbohydrate (48%), protein (25.5%), mucilaginous matter (20%), fat (7.9%), and saponin (4.8%). • Inspite of large potential and high content of protein in fenugreek seeds, however, no reports on molecular structure predictions is available on Trigonella spp. native to this region.

  5. An Introduction to Bioinformatics • Bioinformatics is the application of computer technology for the management of biological information. • Computers are used to gather, store, analyse and integrate biological and genetic information which can then be applied to gene-based sequence determination and structure prediction. • In the present study, Fenugreek (Trigonellafoenum-graecum)protein models were designed and generated using in-silico tools.

  6. NCBI NationalCentre for BiotechnologyInformation (http://www.ncbi.nlm.nih.gov/) • The raw data was collected from NCBI a public domain for the structural analysis. • It is a comprehensive website for biologists including: • biology-related databases, • tools for viewing and analyzing • automated systems for storing and retrieval

  7. NCBI - Home Page (http://www.ncbi.nlm.nih.gov/)

  8. Calculation of Physicochemical properties ProtParam : A tool which allows the computation of various physical and chemical parameters for a given protein . (http://www.expasy.org/tools/Protparam) • Molecular weight • Theoretical PI • Amino acid composition • Atomic composition • Instability index • Aliphatic index and • Grand average of hydropathicity (GRAVY)

  9. The Homepage of ProtParam

  10. Secondary Structure Prediction • GORIV • PSIPRED

  11. GOR IV GOR is information theory based method and it gives the information about Alpha helix, extended strand, and beta turn in the form of propensities.

  12. PSIpred • PSIPRED (bioinf.cs.ucl.ac.uk/psipred) • ALGORITHM It incorporates two feed-forward neural networks which perform an analysis on output obtained from PSI-BLAST   (Position Specific Iterated - BLAST)

  13. Tertiary structure Prediction • Swiss model • Phyre2 • I-TASSER.

  14. Swiss model SWISS MODEL (http://swissmodel.expasy.org) A fully automated protein structure homology-modeling server, accessible via the ExPASy. It is a server used for automated comparative modeling of three-dimensional (3D) protein structures using its sequence.

  15. PHYRE2 Protein Homology/AnalogYRecognition Engine (http://www.sbg.bio.ic.ac.uk/Phyre2/html/page.cgi?id=index) Web-based services for protein structure prediction. Generates reliable protein models when other widely used methods such as PSI-BLAST cannot.

  16. I-TASSER • Server is an Internal service system for protein structure and function predictions. • 3D models was built based on multiple-threading alignments performed by iterative TASSER assembly simulation; functional insights are then derived by matching the predicted models with protein function database imbibed in it .

  17. Result and Discussion

  18. Physicochemical-properties ProtParam computes various physicochemical properties from the protein sequence. The parameters computed by the Server include the molecular weight, theoretical pI, amino acid composition, atomic composition, extinction coefficient, estimated half life, instability index, aliphatic index, and grand average of hydropathicity (GRAVY).

  19. Secondary structure • GOR IV • PSIPRED • Secondary structure of protein was predicted by the formation of alpha helix and ß-sheets. • The results revealed that random coil (69.61%) dominated among secondary structure elements and alpha helices (4.90%) and extended strand (25.49%) were also present

  20. Statistics of Secondary Structure 2D visualization by PSIPred

  21. Tertiary Structure Prediction • Swiss model is homology based method • Automated mode was used to get tertiary structure. But on the basis of structure validation the structure was found to be inappropriate. • Next Phyre2, I-TASSER were used for 3D structure generation. These are threading based software. • Verification : • Through the Structural Analysis and Verification Server (SAVS) the structure by Phyre2 was found to be better as compared to others.

  22. 3D Structure for fenugreek protein

  23. Ramachandran Plot • The Ramachandran plot is a plot of the torsional angles  phi and psi of each residue (amino acids) contained in a peptide.  By making a Ramachandran plot of the protein structure one can determine which torsion angles are lying in allowed region of the plot.

  24. Ramachandran plot by SAVES Server

  25. Conclusion • The predicted protein is stable, globular and basic in nature having pore lining, depicting it as transmembrane protein • The identified protein of fenugreek showed homology with the protein domain of humans and E-coli illustrating that the database available on Apicaeae and Fabaceae family are sparsely populated with sequences. • FFPred predicted its functions in 2 categories: 1) Biological Process: phosphate-containing compound metabolic process with a probability of 95.9% and 2) Molecular function: ATP binding with 98.1% probability.

  26. References • Bukhari, S.B., Bhanger, M.I. and Memon S., (2008). Antioxidative Activity of Extracts from Fenugreek Seeds (Trigonella foenum-graecum) Pak. J. Anal. Environ. Chem.9: 78-83. • Gill S.C. and Von Hippel P.H., (1989). Extinction coefficient. Anal. Biochem.182:319- 328. • Guermeur Y., Geourjon C., Gallinari P. And Deleage G. (1999). Improved performance in protein secondary structure prediction by inhomogeneous score combination. Bioinformatics, 15: 413–21. • Guruprasad K., Reddy B.V.P and Pandit M.W., (1990). Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence. Prot. Eng. 4: 155-164.

  27. Harish, Gupta, A.K., Ram, K., Singh, B., Phulwaria M. and Shekhawat N.S., (2011). Molecular and Biochemical Characterization of Different Accessions of Fenugreek (Trigonella foenum-graecum L.) Libyan Agri. Research Center J. International2: 150-154. • Helambe S.S. and Dande R.P., (2012). Fenugreek (Trigonella foenum-graecum L.): An Overview, IJCPR.2: 169-187 • Ikai A.J., (1980). Thermo stability and aliphatic index of globular proteins. J. Biochem.88: 1895-1898. • Jethra G., Mishra A.K., Pandey P.S. and Chandrasekharan H., (2012) Structure and Function prediction of unknown wheat protein using LOMENTS and I-TASSER. Indian Journal of Agricultural Sciences 82: 867–74. • Zhang Y., (2008). Progress and challenges in protein structure prediction. Current Opinion in Structural Biology,18: 342-348.

  28. GROW MORE FOOD BECAUSE IT IS THE VERY ESSENCE OF OUR LIFE

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