Isolation, characterisation and kinetic studies on SEVA TB ES-31 antigen, a metallo-serine protease of interest in serodiagnosis.

Simple item page
dc.contributor.authorUpadhye, Vijay Jen_US
dc.contributor.authorGomashe, Ashok Ven_US
dc.contributor.authorKumar, Satishen_US
dc.contributor.authorHarinath, Bhaskar Cen_US
dc.date.accessioned2009-01-01en_US
dc.date.accessioned2009-06-01T12:22:22Z
dc.date.available2009-01-01en_US
dc.date.available2009-06-01T12:22:22Z
dc.date.issued2009-01-01en_US
dc.description.abstractBACKGROUND: SEVA TB Excretory secretory-31 (ES-31) antigen, a glycoprotein isolated from M. tb H37Ra culture filtrate, was found to be useful in the serodiagnosis of pulmonary tuberculosis (TB), extrapulmonary TB and in HIV-TB coinfection. Further, it has been shown to be a zinc containing serine protease. AIM: To isolate and purify SEVA TB ES-31 antigen from M. tb H37Ra culture filtrate and study of its enzyme properties and peptide sequence. METHODS: ES-31 antigen was purified from culture filtrate of M. tuberculosis H37Ra strain by ammonium sulphate precipitation, SDS-PAGE fractionation and FPLC. Protease activity of ES-31 antigen was studied using azocasein as substrate. ES-31 antigen was further fractionated by two dimensional polyacrylamide gel electrophoresis (2D PAGE) followed by LCMS-T analysis. RESULTS: Mycobacterial metallo-serine protease was purified 3096 fold from M. tb H37Ra culture filtrate protein. Purified enzyme showed optimum activity at pH 7.0 at 37 degrees C. Of the four substrates explored, the enzyme has shown maximum activity with azocasein and had a Km value of 0.01 mM with specific activity of 6250 x 10(-6) U/mg protein. Further, analysis of ES-31 antigen by 2D PAGE showed two protein spots (A and B). CONCLUSION: Kinetic studies on SEVA TB ES-31 protein, an immunogen with metallo serine protease activity are reported for the first time. Purified enzyme had a Km value of 0.01 mM with azocasein as substrate. Further, study on structure and biological role of serine protease will be of interest.en_US
dc.description.affiliationJamnalal Bajaj Tropical Disease Research Centre, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Wardha.en_US
dc.identifier.citationUpadhye VJ, Gomashe AV, Kumar S, Harinath BC. Isolation, characterisation and kinetic studies on SEVA TB ES-31 antigen, a metallo-serine protease of interest in serodiagnosis. Indian Journal of Tuberculosis. 2009 Jan; 56(1): 22-9en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/110501
dc.language.isoengen_US
dc.source.urihttps://medind.nic.in/ibr/ibrai.shtmlen_US
dc.subject.meshAntigens, Bacterial --chemistry
dc.subject.meshAntigens, Bacterial --isolation & purification
dc.subject.meshAntigens, Bacterial --metabolism
dc.subject.meshBiological Markers --analysis
dc.subject.meshBiological Markers --metabolism
dc.subject.meshCell Culture Techniques
dc.subject.meshElectrophoresis, Gel, Two-Dimensional
dc.subject.meshHumans
dc.subject.meshMass Spectrometry
dc.subject.meshMycobacterium tuberculosis --immunology
dc.subject.meshSerine Endopeptidases --chemistry
dc.subject.meshSerine Endopeptidases --isolation & purification
dc.subject.meshSerine Endopeptidases --metabolism
dc.subject.meshSerologic Tests --methods
dc.subject.meshTuberculosis --diagnosis
dc.subject.meshTuberculosis --immunology
dc.titleIsolation, characterisation and kinetic studies on SEVA TB ES-31 antigen, a metallo-serine protease of interest in serodiagnosis.en_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
Files
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.79 KB
Format:
Plain Text
Description:
Download
Collections
Indian Journal of Tuberculosis