PagP, an integral outer membrane enzyme of e.coli I am working on, has been shown to control the cytoplasmic event of lipopolysaccharide biosynthesis when activated . From the structural details of PagP, it seems that three amino acid residues in the inner leaflet region are organise in the catalytic triad (they have superimopsable image with chymotrypsin catalytic triad). Now, we want to know whether these three residues are actually working as catalytic triad and doing the signal transduction. What reliable experiments can answer this question?
Since, Outer membrane protein is affecting the cytoplasmic event that means there must be some proteins in periplasm and inner membranes that are co-operating in signal transduction. What experiments can allow us to find out which proteins are participating in this putative signal transduction pathway.
Your help will be really appreciated.
Thank you in advance
How to resolve signal transduction pathway in E. coli?