Isolation, characterisation and kinetic studies on SEVA TB ES-31 antigen, a metallo-serine protease of interest in serodiagnosis.
Artigo
em Inglês
| IMSEAR
| ID: sea-110501
ABSTRACT
BACKGROUND:
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.
Texto completo:
DisponíveL
Índice:
IMSEAR (Sudeste Asiático)
Assunto principal:
Espectrometria de Massas
/
Tuberculose
/
Humanos
/
Testes Sorológicos
/
Serina Endopeptidases
/
Biomarcadores
/
Eletroforese em Gel Bidimensional
/
Técnicas de Cultura de Células
/
Mycobacterium tuberculosis
/
Antígenos de Bactérias
Tipo de estudo:
Estudo diagnóstico
Idioma:
Inglês
Ano de publicação:
2009
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS