My laboratory has three major focuses relevant to several human diseases including hypertension, cardiovascular disease, diabetes, autoimmunity and malaria. Our main goal is to define novel points for therapeutic intervention, thus enabling development of new drugs to treat these diseases. To achieve these ends we have adopted a multidisciplinary approach, combining the strengths of cutting edge technologies such as mass spectrometry and chemoproteomics with classical approaches such as physiological studies in isolated tissues, mouse genetics and biochemical methods. In project 1 we utilized a combination of proteomics and muscle physiology to identify ZIPK as a major regulator of Ca2+ independent regulation of smooth muscle contraction. We now believe that ZIPK may be an exquisite drug target for the development of a new generation of antihypertensive drugs. To validate this hypothesis we have generated two knock in flox p ZIPK mouse lines that will enable us to study the consequences of both targeted ZIPK deletion as well as global deletion on cardiovascular physiology as well as during mouse development. In project 2 we utilized a similar approach to define early phosphorylation events in response to activation of PKA and PKG in vivo. Several novel phosphoprotein targets were identified, including CHASM. CHASM shows discrete expression that confines the protein to certain smooth muscles and to type IIa fibers in striated muscle. Studies with the CHASM null mouse shows that the protein plays a critical role in adaptive responses to exercise. CHASM null mice appear to be already exercised adapted, particularly with respect to their cardiovascular responses. This finding suggests that cAMP/cGMP mediated signaling pathways contribute in previously unrecognized ways to smooth and striated muscle phenotype. In project 3 we are utilizing a chemoproteomic approach to define novel antimalarial targets within the Plasmodium falciparum kinome. Plasmodium species have ~90 protein kinases (PfPKs), of these, at least 50 appear to be entirely unique to malaria or at least have no close human orthologs. Because of it uniqueness, the Plasmodium kinome affords an opportunity to develop robust novel antimalarial therapies. In particular we are developing compounds that show broad specificity within the kinome. We believe that single agent drugs targeting two or more malarial protein kinases simultaneously will result in drugs that are inherently resistant to developing drug resistance in the field. We are also using novel inhibitors of PfPKs to define PfPK function in vivo.

Alms GR, Sanz P, Carlson M and Haystead TAJ. (1999) Reg1p targets protein phosphatase 1 to dephosphorylate hexokinase II in Saccharomyces cerevisiae: Characterizing the effects of a phosphatase subunit on the yeast proteome. EMBO J.18 (15):4157-4168.

MacDonald JM, Borman MA, Murányi A, Hartshorne DJ and Haystead TAJ.(2001) Identification of a myosin phosphatase associated kinase regulated by the Rho-kinase pathway. PNAS (98) 2419-2424.

MacDonald, J.A., Eto,M., Borman, M.A., Brautigan,M., and Haystead, T.A.J. (2001). Dual Ser and Thr phosphorylation of CPI-17, an inhibitor of myosin phosphatase, by MYPT-associated kinase. FEBS Lett. 493 (2-3):91-4.

Graves PR, Haystead TA. Molecular biologist's guide to proteomics. Microbiol Mol Biol Rev. 2002 Mar;66(1):39-63; table of contents. Review.

MacDonald JA, Mackey AJ, Pearson WR, Haystead TA. A strategy for the rapid identification of phosphorylation sites in the phosphoproteome. Mol Cell Proteomics. 2002 Apr;1(4):314-22.

Borman MA, MacDonald JA, Muranyi A, Hartshorne DJ, Haystead TA. Smooth muscle myosin phosphatase-associated kinase induces Ca2+ sensitization via myosin phosphatase inhibition. J Biol Chem. 2002 Jun 28;277(26):23441-6.

Graves PR, Kwiek JJ, Fadden P, Ray R, Hardeman K, Coley AM, Foley M, and Haystead TA. Discovery of Novel Targets of Quinoline Drugs in the Human Purine Binding Proteome. Mol Pharmacol December 2002 Dec;62(6):1364-72

Kwiek JJ, Haystead TA, Rudolph J. Kinetic mechanism of quinone oxidoreductase 2 and its inhibition by the antimalarial quinolines.Biochemistry. 2004 Apr 20;43(15):4538-47.

Wooldridge AA, MacDonald JA, Erdodi F, Ma C, Borman MA, Hartshorne DJ, Haystead TA. Smooth muscle phosphatase is regulated in vivo by exclusion of phosphorylation of Threonine 696 of MYPT1 by phosphorylation of Serine 695 in response to cyclic nucleotides. J Biol Chem. 2004 Aug 13;279(33):34496-504.

Graves PR, Winkfield KM, Haystead TA. Regulation of ZIP kinase activity in vitro and in vivo by multi-site phosphorylation.J Biol Chem. 2005 Mar 11;280(10):9363-74.

Kwiek NC, Thacker DF, Datto MB, Megosh HB, Haystead TA. PITK, a PP1 targeting subunit that modulates the phosphorylation of the transcriptional regulator hnRNP K. Cell Signal. 2006 Oct;18(10):1769-78.

Hagerty L, Weitzel DH, Chambers J, Fortner CN, Brush MH, Loiselle D, Hosoya H, Haystead TA. ROCK1 phosphorylates and activates ZIP kinase. J Biol Chem. 2007 Feb 16;282(7):4884-93.

Philip N, Haystead TA. Characterization of a UBC13 kinase in Plasmodium falciparum Proc Natl Acad Sci U S A. 2007 May 8;104(19):7845-50