I have used the baculovirus expression system to over-express the complete intracellular tyrosine kinase domain and a carboxy terminal truncation of the EGF receptor as a soluble protein with the ultimate goal of producing sufficient quantity of protein for crystallographic analysis. The recombinant proteins have been purified to homogeneity by immunoaffinity chromatography and enzymatically characterized in vitro. The activity of the tyrosine kinase domain has been compared to the EGF-activated EGF receptor, and the effect of various activating molecules has been studied.

PUBLICATIONS (resulting from this training, and more recent ones).

Wedegaertner PB, Gill GN. (1989) Activation of the purified protein tyrosine kinase domain of the epidermal growth factor receptor. J Biol Chem. 264:11346-53.

Nishibe S, Wahl MI, Wedegaertner PB, Kim JW, Rhee SG, Carpenter G. (1990) Selectivity of phospholipase C phosphorylation by the epidermal growth factor receptor, the insulin receptor, and their cytoplasmic domains. Proc Natl Acad Sci U S A. 87:424-8.

Wedegaertner PB, Gill GN. (1992) Effect of carboxyl terminal truncation on the tyrosine kinase activity of the epidermal growth factor receptor. Arch Biochem Biophys. 292:273-80.

Wedegaertner PB, Chu DH, Wilson PT, Levis MJ, Bourne HR. (1993) Palmitoylation is required for signaling functions and membrane attachment of Gq alpha and Gs alpha. J Biol Chem. 268:25001-8.

Wedegaertner PB, Bourne HR. (1994) Activation and depalmitoylation of Gs alpha. Cell. 77:1063-70.

Wedegaertner PB, Wilson PT, Bourne HR. (1995) Lipid modifications of trimeric G proteins. J Biol Chem. 270:503-6. Review.

Wedegaertner PB, Bourne HR, von Zastrow M. (1996) Activation-induced subcellular redistribution of Gs alpha. Mol Biol Cell. 7:1225-33.

Iiri T, Backlund PS Jr, Jones TL, Wedegaertner PB, Bourne HR. (1996) Reciprocal regulation of Gs alpha by palmitate and the beta gamma subunit. Proc Natl Acad Sci U S A. 93:14592-7.

Wedegaertner PB. (1998) Lipid modifications and membrane targeting of G alpha. Biol Signals Recept. 7:125-35.

Carman CV, Parent JL, Day PW, Pronin AN, Sternweis PM, Wedegaertner PB, Gilman AG, Benovic JL, Kozasa T. (1999) Selective regulation of Galpha(q/11) by an RGS domain in the G protein-coupled receptor kinase, GRK2. J Biol Chem. 274:34483-92.

Evanko DS, Thiyagarajan MM, Wedegaertner PB. (2000) Interaction with Gbetagamma is required for membrane targeting and palmitoylation of Galpha(s) and Galpha(q). J Biol Chem. 275:1327-36.

Bhattacharyya R, Wedegaertner PB. (2000) Galpha 13 requires palmitoylation for plasma membrane localization, Rho-dependent signaling, and promotion of p115-RhoGEF membrane binding. J Biol Chem. 275:14992-9.

Evanko DS, Thiyagarajan MM, Siderovski DP, Wedegaertner PB. (2001) Gbeta gamma isoforms selectively rescue plasma membrane localization and palmitoylation of mutant Galphas and Galphaq. J Biol Chem. 276:23945-53. Epub 2001 Apr 9.

Wedegaertner PB. (2002) Characterization of subcellular localization and stability of a splice variant of G alpha i2. BMC Cell Biol. 3:12.

Thiyagarajan MM, Bigras E, Van Tol HH, Hebert TE, Evanko DS, Wedegaertner PB. (2002) Activation-induced subcellular redistribution of G alpha(s) is dependent upon its unique N-terminus. Biochemistry. 41:9470-84.

Sterne-Marr R, Tesmer JJ, Day PW, Stracquatanio RP, Cilente JA, O'Connor KE, Pronin AN, Benovic JL, Wedegaertner PB. (2003) G protein-coupled receptor Kinase 2/G alpha q/11 interaction. A novel surface on a regulator of G protein signaling homology domain for binding G alpha subunits. J Biol Chem. 2786050-8.

Bhattacharyya R, Wedegaertner PB. (2003) Characterization of G alpha 13-dependent plasma membrane recruitment of p115RhoGEF. Biochem J. 371:709-20.

Takida S, Wedegaertner PB. (2003) Heterotrimer formation, together with isoprenylation, is required for plasma membrane targeting of Gbetagamma. J Biol Chem. 278:17284-90.

Bhattacharyya R, Wedegaertner PB. (2003) Mutation of an N-terminal acidic-rich region of p115-RhoGEF dissociates alpha13 binding and alpha13-promoted plasma membrane recruitment. FEBS Lett. 540:211-6.

Day PW, Carman CV, Sterne-Marr R, Benovic JL, Wedegaertner PB. (2003) Differential interaction of GRK2 with members of the G alpha q family. Biochemistry. 42:9176-84.

Banerjee J, Wedegaertner PB. (2004) Identification of a novel sequence in PDZ-RhoGEF that mediates interaction with the actin cytoskeleton. Mol Biol Cell. 15:1760-75.

Thiyagarajan MM, Stracquatanio RP, Pronin AN, Evanko DS, Benovic JL, Wedegaertner PB. (2004) A predicted amphipathic helix mediates plasma membrane localization of GRK5. J Biol Chem. 279:17989-95.

Takida S, Wedegaertner PB. (2004) Exocytic pathway-independent plasma membrane targeting of heterotrimeric G proteins. FEBS Lett. 567:209-13.

Day PW, Tesmer JJ, Sterne-Marr R, Freeman LC, Benovic JL, Wedegaertner PB. (2004) Characterization of the GRK2 binding site of Galphaq. J Biol Chem. 279:53643-52. Epub 2004 Oct 7.

Day PW, Wedegaertner PB, Benovic JL. (2004) Analysis of G-protein-coupled receptor kinase RGS homology domains. Methods Enzymol. 390:295-310.

Takida S, Fischer CC, Wedegaertner PB. (2005) Palmitoylation and plasma membrane targeting of RGS7 are promoted by alpha o. Mol Pharmacol. 67:132-9. Epub 2004 Oct 20.

Grabocka E, Wedegaertner PB. (2005) Functional consequences of G alpha 13 mutations that disrupt interaction with p115RhoGEF. Oncogene. 24:2155-65.

Evanko DS, Thiyagarajan MM, Takida S, Wedegaertner PB. (2005) Loss of association between activated Galpha q and Gbetagamma disrupts receptor-dependent and receptor-independent signaling. Cell Signal. 17:1218-28.

Billington CK, Kong KC, Bhattacharyya R, Wedegaertner PB, Panettieri RA Jr, Chan TO, Penn RB. (2005) Cooperative regulation of p70S6 kinase by receptor tyrosine kinases and G protein-coupled receptors augments airway smooth muscle growth. Biochemistry. 44:14595-605.

Escriba PV, Wedegaertner PB, Goni FM, Vogler O. (2007) Lipid-protein interactions in GPCR-associated signaling. Biochim Biophys Acta. 1768:836-52.