Analysis of complex formation and immune response of CFP-10 and ESAT-6 mutants.
ESAT-6 and CFP-10 form a 1:1 heterodimeric complex which contributes to the virulence of Mycobacterium tuberculosis H37Rv. Based on the structure of CFP-10-ESAT-6 complex, we have selected four point mutations each of CFP-10 and ESAT-6 and have analyzed complex formation for the 25 possible combinations between wild-type and mutant CFP-10 and ESAT-6 proteins. We observed that the mutations L25R or F58R of CFP-10 and L29D or L65D of ESAT-6 lead to disruption of complex formation. We have evaluated the immunogenic responses of the wild-type and mutant CFP-10 and ESAT-6 proteins, the wild-type CFP-10-ESAT-6 complex, six complex-forming and two non-complex-forming combinations of wild-type/mutant CFP-10 and ESAT-6 proteins. CFP-10 mutants I21R, L25R, and W43R were found to have better immunogenic potential than wt-CFP-10, while none of the ESAT-6 mutants were better than wt-ESAT-6. Very interestingly, we have discovered that the non-complex-forming mixture of CFP-10-I21R and ESAT-6-L29D gives a strong immunogenic response.
Meher
Akshaya K
AK
Molecular and Structural Biology, Central Drug Research Institute, Lucknow 226001, India.
Lella
Ravi K
RK
Sharma
Charu
C
Arora
Ashish
A
D000936:Antigen-Antibody Complex / Q000276:immunology*; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology* / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology* / Q000494:pharmacology; D049109:Cell Proliferation; D002942:Circular Dichroism; D003001:Cloning, Molecular; D006801:Humans; D009154:Mutation / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000378:metabolism; D011485:Protein Binding; D018417:Th1 Cells / Q000187:drug effects
D000936:Antigen-Antibody Complex; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C412542:CFP 6 protein, Mycobacterium tuberculosis; C093276:ESAT-6 protein, Mycobacterium tuberculosis