The role of N157 of A-ATP synthase subunit B in nucleotide binding and expression, production and structural and mechanistic features of subunits α and ε of Mycobacterium tuberculosis F-ATP synthase

Abstract

NMR and attempts have been invested with crystallisation techniques to garner data to elucidate the structure of this protein. To explore the mechanistic features, chimeric proteins of M. tuberculosis and E. coli (Ec) were generated and have been studied parallel to Ecε and PSε to gain insight into the functioning of Mtε. Mtε on reconstitution with PS3α 3 β 3 γ had no effect on the ATPase activity of the PS3α 3 β 3 γ complex. Ecε also similarly did not alter the activity of the complex PS3α 3 β 3 γ. In contrast, PS3ε on reconstitution increased the ATPase activity of PS3α 3 β 3 γ, highlighting the specific interaction required between the subunits α and ε. This observation was further validated by reconstituting α chi 3 β 3 γ with Mtε that showed increased ATPase activity. Taken together, these studies demonstrate that the shorter length of Mtε is compensated with the longer Mt

Characterization of regulatory central stalk epitopes of mycobacterial- and acetogen F-ATP synthases

“…To engineer the S400C and V525C mutations in the subunit α chi , the plasmid harboring genes that code α chi , γ and β subunits, generated and described in the thesis of Dr. Lavanya Sundararaman (291) was used as a template for the site-directed mutagenesis. The PCR amplification of the entire plasmid was carried out as described in the section 2.2.2 together with Dr. Dhirendra Singh from our group, yielding a single band of 9,316 base pairs, which was visualized on the 1% agarose gel (Figure 3.32).…”