Acta Scientific Veterinary Sciences (ASVS)(ISSN: 2582-3183)

Research Article Volume 2 Issue 3

An In-Silico Structural Characterization of the Buffalo Steroidogenic Proteins

Emmagouni Sharath Kumar Goud, Mamta Pandey, Prashant Singh, Chhama Singh, Vedamurthy Gowdar Veerappa, Dheer Singh, Suneel Kumar Onteru*

Molecular Endocrinology, Functional Genomics, and Systems Biology Lab, Animal Biochemistry Division, ICAR-National Dairy Research Institute, Karnal, India

*Corresponding Author: Suneel Kumar Onteru, Senior Scientist, Molecular Endocrinology, Functional Genomics, and Systems Biology Lab, Animal Biochemistry Division, ICAR-National Dairy Research Institute, Karnal, India.

Received: February 06, 2020; Published: February 21, 2020

×

Abstract

  Post-partum reproductive disorders are of major concern in buffaloes. Mainly these are regulated by steroidogenic proteins, such as CYP17, CYP19, and 3β-HSD. These enzymes are involved in the synthesis of several steroid hormones, an imbalance in the levels of which can lead to the causation of several reproductive illnesses ultimately affecting milk production. In the present in-silico study, we analyzed the structural details of the three steroid enzymes in terms of their physiochemical properties, N- and O-glycosylation sites, phosphorylation sites, secondary structure, surface probability, hydrophilicity and antigenic index. Additionally, the 3D models for the three proteins were enacted in the SWISS-MODEL online tool and the models were assessed by RAMPAGE. It was found that the CYP17 protein sequence of buffalo is small having 247 aminoacids when compared with other species. All the three buffalo steroid proteins were having more than 95% similarity with cattle, sheep and goat, except human. The amino acids responsible for the heme-binding site in CYP17, a catalytic site in CYP19, and 3β-HSD proteins in buffalo were found to be conserved when compared with human protein sequences. The 3D models predicted for the three buffalo steroidogenic proteins were found to be of good quality through Ramachandran plot. Further, the individual phylogenetic trees for each of the protein showed that human proteins are phylogenetically outgroup to buffalo proteins. Overall, the in-silico analysis of the three buffalo steroidogenic proteins could prove to arise new insights for resolving reproductive disorders in the buffaloes.

Keywords: Steroidogenic; In-Silico; Buffalo; Steroid

×

References

  1. http://www.nddb.org/information/stats/milkprodindiaz
  2. http://www.milkproduction.com/Library/Editorial-articles/Milk-quality-in-India/
  3. Puurunen J., et al. “Adrenal androgen production capacity remains high up to menopause in women with polycystic ovary syndrome”. The Journal of Clinical Endocrinology and Metabolism 94.6 (2009): 1973-1978.
  4. Neve EP and Ingelman-Sundberg M. “Cytochrome P450 proteins: retention and distribution from the endoplasmic reticulum”. Current Opinion in Drug Discovery and Development 13.1 (2010): 78-85.
  5. Fernández-Cancio M., et al. “Discordant genotypic sex and phenotype variations in two Spanish siblings with 17α-hydroxylase/17, 20-lyase deficiency carrying the most prevalent mutated CYP17A1 alleles of Brazilian patients”. Sexual Development 11.2 (2017): 70-77.
  6. Kumar OS., et al. “CYP19 (cytochrome P450 aromatase) gene polymorphism in murrah buffalo heifers of different fertility performance”. Research in Veterinary Science 86.3 (2009): 427-437.
  7. Poulos TL., et al. “The 2.6-A crystal structure of Pseudomonas putida cytochrome P-450”. Journal of Biological Chemistry 260.30 (1985): 16122-16130.
  8. Hafez ESE and Hafez B. “Reproduction in farm animals”. Lippincott Williams and Wilkins publications, (7ed) (2000): 261-263.
  9. Chen J., et al. “The correlation of aromatase activity and obesity in women with or without polycystic ovary syndrome”. Journal of Ovarian Research 8.1 (2015): 11.
  10. Kolon TF., et al. “Disorders of sexual development”. Penn Clinical Manual of Urology, Elsevier Health Sciences (2007): 827-852.
  11. Sievers F., et al. “Fast, scalable generation of high‐quality protein multiple sequence alignments using Clustal Omega”. Molecular Systems Biology 7.1 (2011): 539.
  12. http://web.expasy.org/protparam/
  13. Gasteiger, E., et al. “Protein identification and analysis tools on the ExPASy server”. Humana Press (2005): 571-607.
  14. http://www.cbs.dtu.dk/services/NetOGlyc-3.1/
  15. Julenius K., et al. “Prediction, conservation analysis, and structural characterization of mammalian mucin-type O-glycosylation sites”. Glycobiology 15.2 (2005): 153-164.
  16. http://www.cbs.dtu.dk/services/NetNGlyc/ 
  17. Gupta R., et al. “Prediction of N-glycosylation sites in human proteins”. 46 (2004): 203-206.
  18. http://www.cbs.dtu.dk/services/NetPhos/
  19. Blom, N., et al. “Prediction of post‐translational glycosylation and phosphorylation of proteins from the amino acid sequence”. Proteomics 4.6 (2004): 1633-1649.
  20. http://harrier.nagahama-i-bio.ac.jp/sosui/sosuisignal/sosuisignal_submit.html
  21. Gomi M., et al. “High performance system for signal peptide prediction: SOSUI signal”. Journal of chem-bio informatics 4.4 (2004): 142-147.
  22. Garnier J., et al. “GOR method for predicting protein secondary structure from amino acid sequence”. Methods in Enzymology 266.32 (1996): 540-553.
  23. Kyte J and Doolittle RF. “A simple method for displaying the hydropathic character of a protein”. Journal of Molecular Biology 157.1 (1982): 105-132.
  24. Jameson BA and Wolf H. “The antigenic index: a novel algorithm for predicting antigenic determinants”. Bioinformatics 4.1 (1988): 181-186.
  25. https://swissmodel.expasy.org/
  26. Guex N., et al. “Automated comparative protein structure modeling with SWISS‐MODEL and Swiss‐PdbViewer: A historical perspective”. Electrophoresis 30.S1 (2009).
  27. http://mordred.bioc.cam.ac.uk/~rapper/rampage.php
  28. Kumar S., et al. “MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets”. Molecular Biology and Evolution 33.7 (2016): 1870-1874.
  29. Jones DT., et al. “The rapid generation of mutation data matrices from protein sequences”. Bioinformatics 8.3 (1992): 275-282.
  30. Felsenstein J. “Confidence limits on phylogenies: an approach using the bootstrap”. Evolution 39.4 (1985): 783-791.
  31. Hu J., et al. “Cellular cholesterol delivery, intracellular processing and utilization for biosynthesis of steroid hormones”. Nutrition and metabolism 7.1 (2010): 47.
  32. Picado-Leonard., et al. “Cloning and sequence of the human gene for P450cl7 (steroid) 17α-hydroxylase/17, 20 lyase: similarity with the gene for P450c21”. Dna 6.5 (1987): 439-448.
  33. Akhatar MK., et al. “Cytochrome b5 modulation of 17α hydroxylase and 17-20 lyase (CYP17) activities in steroidogenesis”. Journal of Endocrinology 187 (2005): 267-274.
  34. Ghosh D., et al. “X-ray structure of human aromatase reveals an androgen-specific active site”. The Journal of Steroid Biochemistry and Molecular Biology 118.4-5 (2010): 197-202.
  35. Zhao HF., et al. “Molecular cloning, cDNA structure and predicted amino acid sequence of bovine 3β-hydroxy-5-ene steroid dehydrogenase/Δ5-Δ4 isomerase”. FEBS letters 259.1 (1989): 153-157.
  36. Cravioto MDC., et al. “A new inherited variant of the 3β-hydroxysteroid dehydrogenase-isomerase deficiency syndrome: evidence for the existence of two isoenzymes”. The Journal of Clinical Endocrinology and Metabolism 63.2 (1986): 360-367.
  37. Simard J., et al. “Molecular basis of congenital adrenal hyperplasia due to 3 beta-hydroxysteroid dehydrogenase deficiency”. Molecular Endocrinology 7.5 (1993): 716-728.
  38. Pletnev VZ., et al. “Rational Proteomics V: Structure-based mutagenesis has revealed key residues responsible for substrate recognition and catalysis by the dehydrogenase and isomerase activities in human 3β-hydroxysteroid dehydrogenase/isomerase type 1”. The Journal of Steroid Biochemistry and Molecular Biology 101.1 (2006): 50-60.
  39. Guruprasad K., et al. “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”. Protein Engineering, Design, and Selection 4.2 (1990): 155-161.
  40. Weerapana E and Imperiali B. “Asparagine-linked protein glycosylation: from eukaryotic to prokaryotic systems”. Glycobiology 16.6 (2006): 91R-101R.
  41. Mitra N., et al. “N-linked oligosaccharides as outfitters for glycoprotein folding, form and function”. Trends in Biochemical Sciences 31.3 (2006): 156-163.
  42. Shimozawa O., et al. “Core glycosylation of cytochrome P-450 (arom) Evidence for localization of N terminus of microsomal cytochrome P-450 in the lumen”. Journal of Biological Chemistry 268.28 (1993): 21399-21402.
  43. Kaur J and Bose HS. “Passenger Protein Determines Translocation Versus Retention in the Endoplasmic Reticulum for Aromatase Expression”. Molecular Pharmacology 85.2 (2014): 290-300. 
×

Citation

Citation: Suneel Kumar Onteru. “An In-Silico Structural Characterization of the Buffalo Steroidogenic Proteins". Acta Scientific Veterinary Acta Scientific Veterinary Acta Scientific Medical Sciences 2.3 (2020): 01-08.




Member In



News and Events


  • Certification for Review
    Acta Scientific certifies the Editors/reviewers for their review done towards the assigned articles of the respective journals.
  • Submission Timeline for March Issue
    The last date for submission of articles for regular Issues is April 15, 2020.
  • Publication Certificate
    Authors will be issued a "Publication Certificate" as a mark of appreciation for publishing their work.
  • Best Article of the Issue
    The Editors will elect one Best Article after each issue release. The authors of this article will be provided with a certificate of “Best Article of the Issue”.
  • Welcoming Article Submission
    Acta Scientific delightfully welcomes active researchers for submission of articles towards the upcoming issue of respective journals.
  • Contact US