ISSN 1301-109X | e-ISSN 2147-8325
Northwestern Medical Journal
Identification of Hemagglutinin Protein from Streptococcus pneumoniae Pili as a Vaccine Candidate by Proteomic Analysis [Turk J Immunol]
Turk J Immunol. 2018; 6(1): 8-15 | DOI: 10.25002/tji.2018.698

Identification of Hemagglutinin Protein from Streptococcus pneumoniae Pili as a Vaccine Candidate by Proteomic Analysis

Diana Chusna Mufida1, Kusworini Handono2, Sumarno Reto Prawiro3, Sanarto Santoso3
1Faculty of Medicine, University of Jember, Jember, East Java Indonesia, Laboratory of Microbiology, Jember/East Java, Indonesia
2Faculty of Medicine, University Brawijaya, Malang, East Java, Indonesia, Laboratory of Pathology Clinic, Malang/East Java, Indonesia
3Faculty of Medicine, University Brawijaya, Malang, East Java, Indonesia, Laboratory of Microbiology, Malang/East Java, Indonesia

Introduction: Streptococcus pneumoniae is a Gram-positive, bacterial human pathogen that causes a large number of respiratory tract infections, such as pneumonia and sinusitis, as well as invasive diseases such as septicemia and meningitis. Among the surface-exposed virulence factors, the pneumococcal pilus has been shown to play a role in host-cell adhesion. Hemagglutinin activity has been shown to correlate with adhesion ability. The purpose of this study was to identify of S. pneumoniae pili proteins with hemagglutinin activity, and to proteonomically analyze such proteins as potential vaccine candidates. Materials and Methods: The purification of pili was carried out by a pilus cutter method. After SDS-PAGE separation, pili proteins were purified by electroelution and dialysis. After a hemagglutinination assay with mouse erythrocytes, the hemagglutinin protein was identified with Liquid Chromatography/Mass Spectrometry, along with assessments of antigenicity and epitope mapping. Results: SDS-PAGE showed that pili protein had major bands of molecular weights of 67, 54, 25, and 11 kDa. The 54 kDa pili protein was the hemagglutinin. Computational bioinformatics analysis by Mascot Server revealed this hemagglutinin matched with the Rrgb protein of S. pneumoniae (gi: 299856655). Conclusion: The analysis of its antigenicity, and epitope mapping showed that a novel protein was identified as the pilus backbone (Rrgb) of Streptococcus pneumoniae, and has a high antigenicity suitable for further development as a vaccine candidate.

Keywords: Streptococcus pneumoniae, hemagglutinin, pili, proteomic analysis

Identification of Hemagglutinin Protein from Streptococcus pneumoniae Pili as a Vaccine Candidate by Proteomic Analysis

Diana Chusna Mufida1, Kusworini Handono2, Sumarno Reto Prawiro3, Sanarto Santoso3
1Faculty of Medicine, University of Jember, Jember, East Java Indonesia, Laboratory of Microbiology, Jember/East Java, Indonesia
2Faculty of Medicine, University Brawijaya, Malang, East Java, Indonesia, Laboratory of Pathology Clinic, Malang/East Java, Indonesia
3Faculty of Medicine, University Brawijaya, Malang, East Java, Indonesia, Laboratory of Microbiology, Malang/East Java, Indonesia

Giriş: Streptococcus pneumoniae, pnömoni ve sinüzit gibi solunum yolu infeksiyonu olduğu kadar septisemi ve menenjit gibi daha ciddi infeksiyonlara da yol açan Gram-pozitif bir bakteridir. Virulansı belirleyen yüzey ile ilişkili öğeleri içinde, pnömokok pilusu önemli bir rol oynar. Hemaglütünin aktivitesinin ise, bakterinin yapışma özelliği ile ilişkili olduğu gösterilmiştir ve bu proteinlerin proteom ile irdelenmesi, oluşturulabilecek bir aşıda kullanılabilmelerini sağlayabilir. Gereçler ve Yöntemler: Pilusların saflaştırılması, pilusların kesilmesi ile yapılmıştır. Pili proteini SDS-PAGE kullanarak, elektroelusyon ve diyaliz ile saflaştırılmıştır. Fare eritrositleri ile yapılan hemaglütinasyon testinden sonra hemaglütinin proteini, epitop haritalaması ve antijenik özelliği Sıvı Kromatografisi ve Kitle Spektrometresi ile tanımlandı. Bulgular: SDS-PAGE yöntemi ile pili proteinleri ana bantları 67, 52, 25 ve 11kDA moleküler ağırlığında saptandı. 54 KDa ağırlığındaki pilus proteini hemaglütinin antijeni idi. Mascot sunucusu ile yapılan hesaplamalı biyobilgi analizi bu hemaglutininin S. pneumoniae’inin Rrgb proteini ile eşleştiğini gösterdi. Sonuç: Antijen özelliği ve epitop haritalama Streptococcus pneumoniae’dan pilus omurgası (Rrgb) olarak tanımlanan bu yeni proteinin yüksek antijenik özelliğinden dolayı aşı geliştirmede uygun bir aday olabileceğini göstermektedir.

Anahtar Kelimeler: Streptococcus pneumoniae, hemagglutinin, pili, proteomik analiz

Diana Chusna Mufida, Kusworini Handono, Sumarno Reto Prawiro, Sanarto Santoso. Identification of Hemagglutinin Protein from Streptococcus pneumoniae Pili as a Vaccine Candidate by Proteomic Analysis. Turk J Immunol. 2018; 6(1): 8-15
Manuscript Language: English
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