Proteomic analysis of purified protein derivative of Mycobacterium tuberculosis

Clinical Proteomics

Jayasuryan Narayana⁹  Akhilesh Pandey⁸  Anjali Ganjiwale⁹  HC Harsha⁶  Vithal P Myneedu¹  Aditi Chatterjee⁴  Raja Sekhar Nirujogi⁵  Thottethodi Subrahmanya Keshava Prasad²  Anil K Madugundu⁶  Gajanan J Sathe⁴  Renu Verma³  Vinuth N Puttamallesh⁶  Satish Kumar⁷  Jyoti Sharma⁴ 

[1] Department of Microbiology, LRS Institute of TB and RD, New Delhi, IndiaDepartment of Microbiology, LRS Institute of TB and RD, New Delhi, IndiaDepartment of Microbiology, LRS Institute of TB and RD, New Delhi, India;Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaManipal University, Manipal, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaManipal University, Manipal, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaManipal University, Manipal, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaManipal University, Manipal, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaManipal University, Manipal, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaManipal University, Manipal, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, India;Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaMicrotest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaMicrotest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaMicrotest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore, IndiaMicrotest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaSchool of Biotechnology, KIIT University, Bhubaneswar, Odisha, IndiaMicrotest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore, India;Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaManipal University, Manipal, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaManipal University, Manipal, IndiaManipal University, Manipal, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaManipal University, Manipal, India;Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaCentre for Bioinformatics, School of Life Sciences, Pondicherry University, Puducherry, India;Institute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, IndiaInstitute of Bioinformatics, International Tech Park, Whitefield, Bangalore, India;Armed Forces Medical College, Pune, IndiaArmed Forces Medical College, Pune, IndiaArmed Forces Medical College, Pune, India;Johns Hopkins University School of Medicine, Baltimore, USAJohns Hopkins University School of Medicine, Baltimore, USAJohns Hopkins University School of Medicine, Baltimore, USA;Microtest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore, IndiaMicrotest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore, IndiaMicrotest Innovations Pvt. Ltd., International Tech Park, Whitefield, Bangalore, India

关键词: Biomarker;    Broad spectrum antibiotics;    Epitope;    LC-MS/MS;    Mantoux test;   

DOI  :  10.1186/1559-0275-10-8

来源: Humana Press Inc

【 摘 要 】

Abstract

Background

Purified protein derivative (PPD) has been used for more than half a century as an antigen for the diagnosis of tuberculosis infection based on delayed type hypersensitivity. Although designated as “purified,” in reality, the composition of PPD is highly complex and remains ill-defined. In this report, high resolution mass spectrometry was applied to understand the complexity of its constituent components. A comparative proteomic analysis of various PPD preparations and their functional characterization is likely to help in short-listing the relevant antigens required to prepare a less complex and more potent reagent for diagnostic purposes.

Results

Proteomic analysis of Connaught Tuberculin 68 (PPD-CT68), a tuberculin preparation generated from M. tuberculosis, was carried out in this study. PPD-CT68 is the protein component of a commercially available tuberculin preparation, Tubersol, which is used for tuberculin skin testing. Using a high resolution LTQ-Orbitrap Velos mass spectrometer, we identified 265 different proteins. The identified proteins were compared with those identified from PPD M. bovis, PPD M. avium and PPD-S2 from previous mass spectrometry-based studies. In all, 142 proteins were found to be shared between PPD-CT68 and PPD-S2 preparations. Out of the 354 proteins from M. tuberculosis–derived PPDs (i.e. proteins in either PPD-CT68 or PPD-S2), 37 proteins were found to be shared with M. avium PPD and 80 were shared with M. bovis PPD. Alignment of PPD-CT68 proteins with proteins encoded by 24 lung infecting bacteria revealed a number of similar proteins (206 bacterial proteins shared epitopes with 47 PPD-CT68 proteins), which could potentially be involved in causing cross-reactivity. The data have been deposited to the ProteomeXchange with identifier PXD000377.

Conclusions

Proteomic and bioinformatics analysis of different PPD preparations revealed commonly and differentially represented proteins. This information could help in delineating the relevant antigens represented in various PPDs, which could further lead to development of a lesser complex and better defined skin test antigen with a higher specificity and sensitivity.

【 授权许可】

Unknown