| | Name | Initial Conc. (uM) | Volume (fL) | Buffered |
| 1 | Grb2 | 1 | 1000 | No |
| | There is probably a lot of it in the cell: it is also known as Ash (abundant src homology protein). Also Waters et al JBC 271:30 18224 1996 say that only a small fraction of cellular Grb is precipitated out when SoS is precipitated. As most of the Sos seems to be associated with Grb2, it would seem like there is a lot of the latter. Say 1 uM. This would comfortably saturate the SoS. |
| 2 | Sos | 0.1 | 1000 | No |
| | I have tried using low (0.02 uM) initial concs, but these give a very flat response to EGF stim although the overall activation of Ras is not too bad. I am reverting to 0.1 because we expect a sharp initial response, followed by a decline. |
| 3 | Sos* | 0 | 1000 | No |
| | Phosphorylated form of SoS. Nominally this is an inactivation step mediated by MAPK, see Profiri and McCormick 1996 JBC 271(10):5871. I have not put this inactivation in this pathway so this molecule currently only represents a potential interaction point. |
| 4 | Sos*.Grb2 | 0 | 1000 | No |
| | Inactive complex of Sos* with Grb2 due to phosphorylation of the Sos. [Porfiri E and McCormick F. (1996) J Biol Chem. 271(10):5871-5877] |
| 5 | Sos.Grb2 | 0 | 1000 | No |
| | For simplicity I treat the activation of Sos as involving a single complex comprising Sos, Grb2 and Shc*. This is reasonably documented: Sasaoka et al 1994 JBC 269(51):32621-5 Chook et al JBC 1996 271(48):30472 |
