Craig Venter

Hi M Viper, I'm back. So our lab does not focuses on organic synthesis, we basically isolate compunds from microalgae, purify them and define chemical structure (LCMSMS mostly) and biological properties. Lots of analytical chemists and pharmacysts in the lab, but we don't have organic synthesis gurus. So characterization at least won´t be a problem. Do you think that the OH- group resulting from hydrolyzing SO4 on Fig.4 could react to a solid phase with divinyl sulfone or epoxy attached? If not what kind of substitution would you go for if you where in my shoes? Thanks!

Hi thanks for your quick reply and concern. That centered OH- is an issue. I am afraid that if I halogenate the molecule, the uracyl group could end up modified. There is an easier way to get around it. The strain of cyanobacteria also produces a chemical variant of this toxin (see figure 4 ). I am also isolating this deoxy variant, which has very similar toxic effects on mammalian cells. I can use the deoxy variant instead of modifying the cylindrospermopsin. This way I will only have one OH- group to worry about. This toxin inhibits protein synthesis, so we believe it may bind with ribosomes or other proteins related to the translation process. I am not familiarized with pull down assays myself, but I am willing to try different polimers and linkers which might improve my chances. PS: Sorry for the double bond missing on the drawing, you were right.

Hey guys I work with cylindrospemopsin which is a cyanobacterial toxin (see first figure). I want to know how it interacts with mammalian proteins so I wanted to do a “pull down” assay, using the toxin as bait for target proteins. The uracyl group has been proven to be the binding portion to the toxin’s targets. The sulfate group is not involved with the toxic mechanism so I could hydrolyze it with HCl or other acid. This would leave an OH- group to work with. The molecule will have two OH- groups after hydrolysis. I would like to covalently bind this terminal OH- group to a stationary phase (like magnetic beads or spe column) so I can then use the other portion of the toxin as bait for target proteins. Since OH- is not that reactive, maybe I could substitute it for another functional group? What do you recommend?