|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 265, Issue 11, 5990-6001, 04, 1990
MJ Hart, PG Polakis, T Evans and RA Cerione
The abilities of different GTP-binding proteins to serve as
phosphosubstrates for the epidermal growth factor (EGF) receptor/tyrosine
kinase have been examined in reconstituted phospholipid vesicle systems.
During the course of these studies we discovered that a low molecular mass,
high affinity GTP-binding protein from bovine brain (designated as the
22-kDa protein) served as an excellent phosphosubstrate for the
tyrosine-agarose-purified human placental EGF receptor. The EGF-stimulated
phosphorylation of the purified 22-kDa protein occurs on tyrosine residues,
with stoichiometries approaching 2 mol of 32Pi incorporated/mol of
[35S]guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S)-binding sites. The
EGF-stimulated phosphorylation of the brain 22-kDa protein requires its
reconstitution into phospholipid vesicles. No phosphorylation of this
GTP-binding protein is detected if it is simply mixed with the purified EGF
receptor in detergent solution or if detergent is added back to lipid
vesicles containing the EGF receptor and the 22-kDa protein. The
EGF-stimulated phosphorylation of this GTP-binding protein is also markedly
attenuated by guanine nucleotides, i.e. GTP, GTP gamma S, or GDP,
suggesting that maximal phosphorylation occurs when the GTP- binding
protein is in a guanine nucleotide-depleted state. Purified preparations of
the 22-kDa phosphosubstrate do not cross-react with antibodies against the
ras proteins. However, they do cross-react against two different peptide
antibodies generated against specific sequences of the human platelet (and
placental) GTP-binding protein originally designated Gp (Evans, T., Brown,
M. L., Fraser, E. D., and Northrup, J. K. (1986) J. Biol. Chem. 261,
7052-7059) and more recently named G25K (Polakis, P. G., Synderman, R., and
Evans, T. (1989) Biochem. Biophys. Res. Commun. 160, 25-32). When highly
purified preparations of the human platelet Gp (G25K) protein are
reconstituted with the purified EGF receptor into phospholipid vesicles, an
EGF- stimulated phosphorylation of the platelet GTP-binding protein occurs
with a stoichiometry approaching 2 mol of 32Pi incorporated/mol of [35S]GTP
gamma S-binding sites. As is the case for the brain 22-kDa protein, the
EGF-stimulated phosphorylation of the platelet GTP-binding protein is
attenuated by guanine nucleotides. Overall, these results suggest that the
brain 22-kDa phosphosubstrate for the EGF receptor is very similar, if not
identical, to the Gp (G25K) protein. Although guanine nucleotide binding to
the brain 22-kDa protein or to the platelet. GTP-binding protein inhibits
phosphorylation, the phosphorylated GTP-binding proteins appear to bind
[35S]GTP gamma S slightly better than their nonphosphorylated counterparts.
The identification and characterization of an epidermal growth factor- stimulated phosphorylation of a specific low molecular weight GTP- binding protein in a reconstituted phospholipid vesicle system
Department of Biochemistry, Cell and Molecular Biology, Cornell University, Ithaca, New York 14853.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  S. Tu, W. J. Wu, J. Wang, and R. A. Cerione Epidermal Growth Factor-dependent Regulation of Cdc42 Is Mediated by the Src Tyrosine Kinase J. Biol. Chem., December 5, 2003; 278(49): 49293 - 49300. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. C. Bock, P. O. Vacratsis, E. Qamirani, and K. A. Gallo Cdc42-induced Activation of the Mixed-Lineage Kinase SPRK in Vivo. REQUIREMENT OF THE Cdc42/Rac INTERACTIVE BINDING MOTIF AND CHANGES IN PHOSPHORYLATION J. Biol. Chem., May 5, 2000; 275(19): 14231 - 14241. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. I. Johnson Cdc42: An Essential Rho-Type GTPase Controlling Eukaryotic Cell Polarity Microbiol. Mol. Biol. Rev., March 1, 1999; 63(1): 54 - 105. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. J. McCallum, J. W. Erickson, and R. A. Cerione Characterization of the Association of the Actin-binding Protein, IQGAP, and Activated Cdc42 with Golgi Membranes J. Biol. Chem., August 28, 1998; 273(35): 22537 - 22544. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. E. Ullian, J. R. Raymond, M. C. Willingham, and R. V. Paul Regulation of vascular angiotensin II receptors by EGF Am J Physiol Cell Physiol, October 1, 1997; 273(4): C1241 - C1249. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Hess, A. H. Ross, R.-G. Qiu, M. Symons, and J. H. Exton Role of Rho Family Proteins in Phospholipase D Activation by Growth Factors J. Biol. Chem., January 17, 1997; 272(3): 1615 - 1620. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Poppleton, H. Sun, D. Fulgham, P. Bertics, and T. B. Patel Activation of G[IMAGE] by the Epidermal Growth Factor Receptor Involves Phosphorylation J. Biol. Chem., March 22, 1996; 271(12): 6947 - 6951. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. M. Graves, K. E. Bornfeldt, J. S. Sidhu, G. M. Argast, E. W. Raines, R. Ross, C. C. Leslie, and E. G. Krebs Platelet-derived Growth Factor Stimulates Protein Kinase A through a Mitogen-activated Protein Kinase-dependent Pathway in Human Arterial Smooth Muscle Cells J. Biol. Chem., January 5, 1996; 271(1): 505 - 511. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Dash, M. Aepfelbacher, and W. Siess Integrin [IMAGE][IMAGE][IMAGE][IMAGE]-mediated Translocation of CDC42Hs to the Cytoskeleton in Stimulated Human Platelets J. Biol. Chem., July 21, 1995; 270(29): 17321 - 17326. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |