You can check the mRNA level of the lead treated staph, and compare it with the WT. Then pay attention to these over expression or low expression genes, you may find some targets. And then try to get the gene mutant strain or over express this gene, check whether the staph has any difference with Lead treatment.

But It's not easy to get the result you want, because the lead may make its function not through interacting with some proteins rather than breaking the cell or some other effect.

Good luck.

Lin

A common approach is to use the compound to select for resistant mutant strains, then compare the genomes of the resistant strains to that of the parent. The mutated genes are potential targets, or may suggest the target based on their functions.. This method is not always successful, however, because sometimes  mutations occur in genes other than the target, and with no obvious relationship to a target. The next step is to select for even higher level resistance using the mutant strain. This 2-step procedure may uncover the target.

Hi James, your lead molecule is probably binding to free thiols (SH) in Staph proteins. Pb-S compounds are very insoluble.

You could block free SH groups in Staph by using maleimide or fluorescent maleimide compounds, such as:

http://www.sigmaaldrich.com/catalog/product/sigma/38132?lang=en®ion=US

If  the maleimide mimics the effect of your lead molecule, then it is probably acting through free SH in Staph.

If the maleimide does not inhibit Staph, but addition of your lead molecule to the maleimide treated proteins does, then things will get interesting.

Interesting. I thought James was talking about a lead (leed) molecule as in a drug discovery program. It didn't occur to me he might have been talking about the metal lead (led).

I would think treating bacteria with a maleimide, if it gets inside the cells, would be toxic due to reacting with all the thiols in the cells. It might even be toxic from the outside if it inhibits essential membrane protein, such as in the respiratory chain.

Hi

I am really very sorry to con fuse you all. It is a lead compound (i.e. a "leading" compound, not lead metal), that i have extracted from natural source and found it being active on staph aureus and inhibiting the growth. Thus i would like to see protein profile in comparison to control and treated.

what would be bet easy way to look in to  , i have tried 2D and finding very difficult to standardizing it

Probably some useful points will be in attached article.

Article Comparative study of Surface Plasmon Resonance, electrochemi...