Investigation of the membrane interactions of Mycobacterium tuberculosis secreted factors ESAT-6 and CFP-10
Mycobacterium tuberculosis is the causative agent of the infectious disease tuberculosis, infecting about one-third of human race. Infections are not vast in the United States when compared to many other countries, yet many cases seen in the United States are of those who have immigrated to the United States. The Hispanic/Latino population in the United States contributes to a large fraction of those infected with tuberculosis in the United States. During this study we purified recombinant Mycobacterium tuberculosis secreted factors ESAT-6 and CFP-10 from Escherichia coli. Here we have introduced a method in which we can refold and purify insoluble ESAT-6 using one column. Our purified proteins when incubated together form a heterocomplex just as previous works have shown. We have subjected each protein, ESAT-6, CFP-10, and the heterocomplex to a K+ release assay in which we use prepared liposomes filled with K+ to show protein activity. We have shown that our recombinant proteins are capable of membrane disruption at a low pH. The work presented here builds a base for future biochemical studies in protein-membrane interaction and providing information on how such reactions may contribute to latency and reactivation of Mycobacterium tuberculosis infection.
De Leon, Joaquin A., "Investigation of the membrane interactions of Mycobacterium tuberculosis secreted factors ESAT-6 and CFP-10" (2012). ETD Collection for University of Texas, El Paso. AAI1518195.