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Triton X‐114 Fractionated Subcellular Proteome of Leptospira interrogans Shows Selective Enrichment of Pathogenic and Outer Membrane Proteins in the Detergent Fraction
Author(s) -
Thoduvayil Sikha,
Dhandapani Gunasekaran,
Brahma Rahul,
Devasahayam Arokia Balaya Rex,
Mangalaparthi Kiran K.,
Patel Krishna,
Kumar Manish,
Tennyson Jebasingh,
Satheeshkumar P. K.,
Kulkarni Mahesh J.,
Pinto Sneha M.,
Prasad T. S. Keshava,
Madanan Madathiparambil G.
Publication year - 2020
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.202000170
Subject(s) - proteome , bacterial outer membrane , membrane protein , pathogenic bacteria , chemistry , cell fractionation , leptospira interrogans , biology , leptospira , chromatography , proteomics , biochemistry , membrane , bacteria , microbiology and biotechnology , escherichia coli , gene , serotype , genetics
The Triton X‐114‐based solubilization and temperature‐dependent phase separation of proteins is used for subcellular fractionation where, aqueous, detergent, and pellet fractions represents cytoplasmic, outer membrane (OM), and inner membrane proteins, respectively. Mass spectrometry‐based proteomic analysis of Triton X‐114 fractions of proteomic analysis of Leptospira interrogans identified 2957 unique proteins distributed across the fractions. The results are compared with bioinformatics predictions on their subcellular localization and pathogenic nature. Analysis of the distribution of proteins across the Triton X‐114 fractions with the predicted characteristics is performed based on “number” of unique type of proteins, and “quantity” which represents the amount of unique protein. The highest number of predicted outer membrane proteins (OMPs) and pathogenic proteins are found in aqueous and pellet fractions, whereas detergent fraction representing the OM has the highest quantity of OMPs and pathogenic proteins though lower in number than the aqueous and pellet fractions. This leaves the possibility of an upsurge in pathogenic proteins and OMPs on the OM under pathogenic conditions suggesting their potential use to combat leptospirosis. Further, the Triton X‐114 subcellular fractions are more correlated to enrichment of pathogenic proteins predicted by MP3 software than predicted localization.