Biology Reference
In-Depth Information
9.
Microtubules-stabilizing buffer: 80 mM PIPES/KOH, 1 mM MgCl
2
,1mM
EGTA, 30% (v/v) glycerol, and 1 mM GTP, pH 6.8
10.
Extraction buffer: 100 mM PIPES, 1 mM MgSO
4
, 2 mM EGTA, 100
m
M
EDTA, and 2 M glycerol, pH 6.75
11.
Stripping buffer: 625 mMTris-HCl, 100 mM
b
-mercaptoethanol, and 2% (w/v)
SDS, pH 6.7
CONCLUDING COMMENTS
Activated Gs
a
, translocated to the cytosol from the plasma membrane, appears to
directly interact with tubulin and acts as a direct modulator of microtubule dynamics,
due primarily to the activation of GTPase by Gs
a
. This provides a mechanism for the
modulation of microtubule-based cellular structures by the activation of G-protein-
coupled receptors and does so in a manner independent of second messenger gener-
ation. Hopefully, this chapter has delineated a number of methods that will allow
further elucidation of this underexplored aspect of cell biology.
Acknowledgments
Much of the science described in this chapter was supported by NIH (MH 39595, DA020568,
MH 07800) and the Veteran's Administration (BX11049).
References
