protein interaction study - (Aug/06/2006 )

hello all

i am interested in protein protein interactions.... i mean to study the effect of certain candidate on another type of protein...
how to start? i thought of studying the effect of protein 1 on the phosphorylation state of protein 2
what else do u suggest?
thankx

-spanishflower-

QUOTE (spanishflower @ Aug 7 2006, 04:30 PM)

I assume you know if they actually interact. Have you done IP? Yeast two-hybrid? Gel filtration with and without light scattering? Isothermal titration calorimetry? SPR?

-swanny-

hi swanny
no i dont know if they are actually reacting as i was checking the effect of this protein 1 on ( protein 2 + protein 3 which i know that they bind together)
now i want to study wether protein react with protein 1 or 3

-spanishflower-

Seeing as how you're not certain which protein is the interacting partner, I think you'd need to try something where proteins 2 and 3 are separated, such as yeast two-hybrid, so you can find an individual partner (unless the interaction is at the dimer face...).
All of this is hypothetical until you know there is an interaction with the complex. Perhaps a GST pulldown could answer that problem, then if the interaction is real, you could test each of the partners in turn.

-swanny-