Structural and Functional Characterization of a Hypothetical Protein from Streptococcus mitis Using In silico Approaches

Md. Easin Mia

Department of Biotechnology and Genetic Engineering, Noakhali Science and Technology University, Noakhali-3814, Bangladesh.

Tanvir Hossain Emon

Department of Biotechnology and Genetic Engineering, Noakhali Science and Technology University, Noakhali-3814, Bangladesh.

Md. Shipul Islam

Department of Biotechnology and Genetic Engineering, Noakhali Science and Technology University, Noakhali-3814, Bangladesh.

Md. Anwar Hossain *

Department of Biotechnology and Genetic Engineering, Noakhali Science and Technology University, Noakhali-3814, Bangladesh.

*Author to whom correspondence should be addressed.


Abstract

Streptococcus mitis, a member of the Viridans Group Streptococci (VGS), is commonly found in the normal flora of the human oropharynx, and can cause various infections in humans, including orbital cellulitis, infective endocarditis, and bacteremia in neutropenic individuals. Identifying specific proteins, particularly hypothetical proteins (HPs), is crucial for developing effective treatments. This research aimed to determine the structure as well as the function of a nonannotated hypothetical protein (HP) from Streptococcus mitis. Several aspects of the HP (accession no. BCJ11593.1), including physiochemical properties, 3D structures, and functional annotation, were predicted using a variety of computational tools, followed by further validation and quality assessment.  The HP was identified as a cytoplasmic and stable protein. NCBI-CD search, and InterProScan, two functional annotation tools, predicted that the target HP was the ribosome-associated protein YbcJ, specifically the S4-like RNA-binding protein. Secondary structural investigations found that the alpha helix was prevalent. The study determined its three-dimensional (3D) structure through homology modeling by the SWISS-MODEL server and verified it using quality evaluation tools such as PROCHECK, QMEAN, ERRAT, and ProSA. The findings lay the groundwork for potential antibacterial treatments, highlighting the importance of understanding the structure and function of specific proteins in Streptococcus mitis.

Keywords: Functional annotation, hypothetical protein, in silico characterization, Streptococcus mitis, three-dimensional structure


How to Cite

Mia , Md. Easin, Tanvir Hossain Emon, Md. Shipul Islam, and Md. Anwar Hossain. 2023. “Structural and Functional Characterization of a Hypothetical Protein from Streptococcus Mitis Using In Silico Approaches”. Asian Journal of Biotechnology and Genetic Engineering 6 (2):236-48. https://www.journalajbge.com/index.php/AJBGE/article/view/117.

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References

Choi HP, Juarez S, Ciordia S, Fernandez M, Bargiela R, Albar JP, Mazumdar V, Anton BP, Kasif S, Ferrer M, Steffen M. Biochemical characterization of hypothetical proteins from Helicobacter pylori. PLoS One. 2013;8(6):e66605.

Morozova O, Marra MA. Applications of next-generation sequencing technologies in functional genomics. Genomics. 2008; 92(5):255-64.

Shahbaaz M, Bisetty K, Ahmad F, Hassan I. Current advances in the identification and characterization of putative drug and vaccine targets in the bacterial genomes. Current Topics in Medicinal Chemistry. 2016;16(9):1040-69.

Nimrod G, Schushan M, Steinberg DM, Ben-Tal N. Detection of functionally important regions in “hypothetical proteins” of known structure. Structure. 2008;16(12): 1755-63.

Pellegrini M, Marcotte EM, Thompson MJ, Eisenberg D, Yeates TO. Assigning protein functions by comparative genome analysis: Protein phylogenetic profiles. Proceedings of the National Academy of Sciences. 1999;96(8):4285-8.

Idrees S, Nadeem S, Kanwal S, Ehsan B, Yousaf A, Nadeem S, Rajoka MI. In silico sequence analysis, homology modeling, and function annotation of Ocimum basilicum hypothetical protein G1CT28_OCIBA. International Journal Bioautomation. 2012;16(2):111.

Lubec G, Afjehi-Sadat L, Yang JW, John JP. Searching for hypothetical proteins: theory and practice based upon original data and literature. Progress in Neurobiology. 2005;77(1-2):90-127.

Naqvi AA, Rahman S, Zeya F, Kumar K, Choudhary H, Jamal MS, Kim J, Hassan MI. Genome analysis of Chlamydia trachomatis for functional characterization of hypothetical proteins to discover novel drug targets. International Journal of Biological Macromolecules. 2017;96:234-40.

Naqvi AA, Anjum F, Khan FI, Islam A, Ahmad F, Hassan MI. Sequence analysis of hypothetical proteins from Helicobacter pylori 26695 to identify potential virulence factors. Genomics Inform. 2016;14(3):125-35.

Yang Z, Zeng X, Tsui SK. Investigating function roles of hypothetical proteins encoded by the Mycobacterium tuberculosis H37Rv genome. BMC Genomics. 2019;20:1-0.

Islam MS, Shahik SM, Sohel M, Patwary NI, Hasan MA. In silico structural and functional annotation of hypothetical proteins of Vibrio cholerae O139. Genomics & Informatics. 2015;13(2):53.

Facklam R. What happened to the streptococci: overview of taxonomic and nomenclature changes? Clinical Microbiology Reviews. 2002;15(4):613-30.

Douglas CW, Heath J, Hampton KK, Preston FE. Identity of viridans streptococci isolated from cases of infective endocarditis. Journal of Medical Microbiology. 1993;39(3):179-82.

Jaing TH, Chiu CH, Hung IJ. Successful treatment of meningitis caused by highly-penicillin-resistant Streptococcus mitis in a leukemic child. Chang Gung Medical Journal. 2002;25(3):190-3.

Colomba C, Garbo V, Boncori G, Albano C, Bagarello S, Condemi A, Giordano S, Canduscio LA, Gallo C, Parrinello G, Cascio A. Streptococcus mitis as a New Emerging Pathogen in Pediatric Age: Case Report and Systematic Review. Antibiotics. 2023;12(7):1222.

Engen SA, Rørvik GH, Schreurs O, Blix IJ, Schenck K. The oral commensal Streptococcus mitis activates the aryl hydrocarbon receptor in human oral epithelial cells. International Journal of Oral Science. 2017;9(3):145-50.

Zheng W, Tan TK, Paterson IC, Mutha NV, Siow CC, Tan SY, Old LA, Jakubovics NS, Choo SW. Strepto Base: an oral Streptococcus mitis group genomic resource and analysis platform. PloS One. 2016;11(5):e0151908.

Almatrafi MA, Almuflihi A, Jarwann R, Alghraibi S, Ashgar E, Alsahaf N, Aburziza AJ. Viridans group Streptococcus meningitis in an immunocompetent child: A case report. The American Journal of Case Reports. 2021;22:e933089-1.

Yiş R, Yüksel CN, Derundere U, Yiş U. Meningitis and white matter lesions due to Streptococcus mitis in a previously healthy child. Mikrobiyoloji Bulteni. 2011;45(4):741-5.

Kutlu SS, Sacar S, Cevahir N, Turgut H. Community-acquired Streptococcus mitis meningitis: A case report. International Journal of Infectious Diseases. 2008;12(6):e107-9.

Curtis H, Dirk G, Rob K, Sahar A, Badger JH, Chinwalla AT, Creasy HH, Earl AM, Fitzgerald MG, Fulton RS, Giglio MG. Structure, function, and diversity of the healthy human microbiome. Nature. 2012;486:207-14.

Shelburne SA, Sahasrabhojane P, Saldana M, Yao H, Su X, Horstmann N, Thompson E, Flores AR. Streptococcus mitis strains causing severe clinical disease in cancer patients. Emerging Infectious Diseases. 2014;20(5):762.

Fukayama H, Shoji K, Yoshida M, Iijima H, Maekawa T, Ishiguro A, Miyairi I. Bacterial meningitis due to the Streptococcus mitis group in children with cerebrospinal fluid leak. IDCases. 2022;27:e01406.

Tunkel AR, Sepkowitz KA. Infections caused by Viridans streptococci in patients with neutropenia. Clinical Infectious Diseases. 2002;34(11):1524-9.

Chaemsaithong P, Lertrut W, Kamlungkuea T, Santanirand P, Singsaneh A, Jaovisidha A, Pakdeeto S, Mongkolsuk P, Pongchaikul P. Maternal septicemia caused by Streptococcus mitis: A possible link between intra-amniotic infection and periodontitis. Case report and literature review. BMC Infectious Diseases. 2022; 22(1):562.

Balkundi DR, Murray DL, Patterson MJ, Gera R, Scott-Emuakpor A, Kulkarni R. Penicillin-resistant Streptococcus mitis as a cause of septicemia with meningitis in febrile neutropenic children. Journal Pediatric Hematology/Oncology. 1997; 19(1):82-5.

Benson DA, Karsch-Mizrachi I, Lipman DJ, Ostell J, Rapp BA, Wheeler DL. GenBank. Nucleic Acids Research. 2002;30(1):17-20.

Gasteiger E, Gattiker A, Hoogland C, Ivanyi I, Appel RD, Bairoch A. ExPASy: The proteomics server for in-depth protein knowledge and analysis. Nucleic Acids Research. 2003;31(13):3784- 8.

Yu CS, Chen YC, Lu CH, Hwang JK. Prediction of protein subcellular localization. Proteins: Structure, Function, and Bioinformatics. 2006;64(3):643-51.

Yu NY, Wagner JR, Laird MR, Melli G, Rey S, Lo R, Dao P, Sahinalp SC, Ester M, Foster LJ, Brinkman FS. PSORTb 3.0: improved protein subcellular localization prediction with refined localization subcategories and predictive capabilities for all prokaryotes. Bioinformatics. 2010; 26(13):1608-15.

Wu CH, Huang H, Yeh LS, Barker WC. Protein family classification and functional annotation. Computational Biology and Chemistry. 2003;27(1):37-47.

Lu S, Wang J, Chitsaz F, Derbyshire MK, Geer RC, Gonzales NR, Gwadz M, Hurwitz DI, Marchler GH, Song JS, Thanki N. CDD/SPARCLE: the conserved domain database in 2020. Nucleic Acids Research. 2020;48(D1):D265-8.

Apweiler R, Attwood TK, Bairoch A, Bateman A, Birney E, Biswas M, Bucher P, Cerutti L, Corpet F, Croning MD, Durbin R. The InterPro database, an integrated documentation resource for protein families, domains and functional sites. Nucleic Acids Research. 2001;29(1):37-40.

Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. Journal of Molecular Biology. 1990; 215(3):403-10.

Combet C, Blanchet C, Geourjon C, Deleage G. NPS@: network protein sequence analysis. Trends in Biochemical Sciences. 2000;25(3):147-50.

Buchan DW, Jones DT. The PSIPRED protein analysis workbench: 20 years on. Nucleic Acids Research. 2019;47(W1): W402-7.

Waterhouse A, Bertoni M, Bienert S, Studer G, Tauriello G, Gumienny R, Heer FT, De Beer TA, Rempfer C, Bordoli L, Lepore R. SWISS-MODEL: Homology modeling of protein structures and complexes. Nucleic Acids Research. 2018; 46(W1):W296-303.

Laskowski RA, MacArthur MW, Moss DS, Thornton JM. PROCHECK: a program to check the stereochemical quality of protein structures. Journal of Applied Crystallography. 1993;26(2):283-91.

Colovos C, Yeates TO. Verification of protein structures: Patterns of nonbonded atomic interactions. Protein Science. 1993;2(9):1511-9.

Benkert P, Biasini M, Schwede T. Toward the estimation of the absolute quality of individual protein structure models. Bioinformatics. 2011;27(3):343-50.

Wiederstein M, Sippl MJ. ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Research. 2007; 35(suppl_2):W407-10.

Barragán-Osorio L, Giraldo G, J Almeciga-Diaz C, Aliev G, E Barreto G, Gonzalez J. Computational analysis and functional prediction of ubiquitin hypothetical protein: a possible target in Parkinson disease. Central Nervous System Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry-Central Nervous System Agents). 2016;16(1):4-11.

Sen T, Verma NK. Functional annotation and curation of hypothetical proteins present in a newly emerged serotype 1c of Shigella flexneri: Emphasis on selecting targets for virulence and vaccine design studies. Genes. 2020;11(3):340.

Gopalakrishna S, Pearce SF, Dinan AM, Rosenberger FA, Cipullo M, Spåhr H, Khawaja A, Maffezzini C, Freyer C, Wredenberg A, Atanassov I. C6orf203 is an RNA-binding protein involved in mitochondrial protein synthesis. Nucleic Acids Research. 2019;47(17):9386-99.

Carter AP, Clemons WM, Brodersen DE, Morgan-Warren RJ, Wimberly BT, Ramakrishnan V. Functional insights from the structure of the 30S ribosomal subunit and its interactions with antibiotics. Nature. 2000;407(6802):340-8.

Volpon L, Lievre C, Osborne MJ, Gandhi S, Iannuzzi P, Larocque R, Cygler M, Gehring K, Ekiel I. The solution structure of YbcJ from Escherichia coli reveals a recently discovered αL motif involved in RNA binding. Journal of Bacteriology. 2003;185(14):4204-10.

Stenum TS, Kumar AD, Sandbaumhüter FA, Kjellin J, Jerlström-Hultqvist J, Andrén PE, Koskiniemi S, Jansson ET, Holmqvist E. RNA interactome capture in Escherichia coli globally identifies RNA-binding proteins. Nucleic Acids Research. 2023;22; 51(9):4572-87.

Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped blast and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Research. 1997; 25(17):3389-402.

Ngernyuang N, Yan W, Schwartz LM, Oh D, Liu YB, Chen H, Shao R. A heparin binding motif rich in arginine and lysine is the functional domain of YKL-40. Neoplasia. 2018;20(2):182-92.