Owen Nadeau, Research Assistant Professor

Department of Biochemistry and Molecular Biology

University of Kansas Medical Center

913-588-3486
onadeau@kumc.edu

---

Education & Experience

• Indiana University of Pennsylvania, Indiana, PA, B.S., Chemistry, 1981
• Indiana University of Pennsylvania, Indiana, PA M.S., Chemistry, 1984
• University of Vermont, Burlington, VT, Ph.D., Biochemistry, 1991
• University of Tennessee, Memphis, TN, Postdoctoral Fellow (Biochemistry) 1991-1997
• University of Tennessee, Memphis, TN, Postdoctoral Fellow (Pathology) 1997
• Hexos Inc., Bothell, WA, Senior Scientist 1997-1999
• University of Missouri-Kansas City, Research Assistant Professor, School of Biological Sciences1999-2002
• University of Kansas Medical Center, Research Assistant Professor, Department of Biochemistry and Molecular Biology, 2003-present

Representative Publications
click here for PubMed listings

Major Research Interest

Our primary goal and continuing efforts are centered on determining how subunit interactions affect the activation state of phosphorylase kinase (PhK), a 1.3 MDa hexadecameric complex comprising four copies of four distinct subunits, termed α, β, γ and δ, which is endogenous calmodulin. In the PhK complex, the catalytic γ subunit is activated by neural (Ca2+), hormonal (cAMP and Ca2+) and metabolic (ADP) stimuli, which are integrated through allosteric sites on the regulatory α, β and δ subunits. This activation of PhK by diverse physiological signals allows for tight control of glycogenolysis. Activation of PhK by Ca2+ in skeletal muscle directly links muscle contraction with energy production in the cascade activation of glycogen utilization.

To analyze the subunit interactions of PhK we have developed a data-based approach to detect protein-protein interactions using a combination of chemical cross-linking and mass spectrometric (MS) methodologies. Recent developments in MS technologies have significantly reduced the amount of starting material required to detect specific contact regions between interacting proteins, leading to a resurgence in classical protein chemical approaches, including chemical cross-linking. Despite the resolving power of MS, considerable computational analyses of digests from conjugates of interacting proteins are required to predict all the possible masses that can arise from monoderivatization, intramolecular cross-linking, incomplete digestion of the target proteins, hydrolysis and other side reactions associated with chemical cross-linking. Considering the complexity of the products formed, we have developed a search engine that predicts both conjugates and side-products of such reactions. The search engine automatically eliminates false positives arising from known competing side reactions, thus allowing for rapid screening of MS peptide maps of protein conjugates for potential cross-linked peptides, which are then ranked hierarchically by theoretical best match. Since several combinations of cross-linking can lead to identical mass assignments within the error limits of state-of-the-art MS technologies, all cross-linked peptides must be verified by fragmentation (MS/MS) analyses, which in turn generate even more complex forms of data output that often must be annotated by hand. To eliminate this ‘second’ rate limiting step in the method, we are currently expanding the search engine’s capability to generate potential fragment ion patterns for those cross-linked peptides that are predicted in the first round of the analysis. A web-based version of the engine is under construction, and the initial results can be viewed at http://funnybase.umkc.edu/prot_cross3/.

Representative Publications

Owen W. Nadeau, David W. Anderson, Qing Yang, Antonio Artigues, Justin E. Paschall, Gerald J. Wyckoff, Jennifer L. McClintock and Gerald M. Carlson (2007) “Evidence for the Location of the Allosteric Activation Switch in the Multisubunit Phosphorylase Kinase Complex from Mass Spectrometric Identification of Chemically Cross-linked Peptides.” J. Mol. Biol. 365: 1429-1445.

Nadeau, O.W. (2006) “Protein-protein Interaction Analysis: Chemical Cross-linking,” in Encyclopedic Reference of Genomics and Proteomics in Molecular Medicine, Eds. K. Ruckpaul and D. Ganten, Springer-Verlag, Heidelberg, online version at http://www.springerlink.com/content/n4425664628508x7/fulltext.html, 2006

Priddy, T.S., MacDonald. B.A., Heller, W.T., Nadeau, O.W., Trewhella, J., and Gerald M. Carlson, (2005) Ca2+-induced Structural Changes in Phosphorylase Kinase Detected by Small-angle X-ray Scattering.” Protein Sci.
14: 1039-1048.

Marshall, S, Nadeau, O.W. and Yamasaki, K, (2005) "Glucosamine-induced Activation of Glycogen Biosynthesis in Isolated Adipocytes: Evidence for a Rapid Allosteric Control Mechanism within the Hexoseamine Biosynthesis Pathway, J. Biol. Chem. 280: 11018-11024.

Nadeau, O.W., Gogol E. and Carlson, G. M. (2005) “Cryoelectron Microscopy Reveals New Features in the Three-dimensional Structure of Phosphorylase Kinase". Protein Sci. 14: 914-920.

Nadeau, O.W. and Carlson, G.M. (2005) “Protein Interactions Captured by Chemical Cross-linking,” in Protein-Protein Interactions, A Molecular Cloning Manual, 2nd edition, Chap. 7, Eds. E. Golemis and P. Adams, Cold Spring Harbor, New York, pp 105-127.

Marshal, S, Nadeau, O.W. and Yamasaki, K. (2004) "Dynamic Actions of Glucose and Glucosamine on Hexosamine Biosynthesis in Isolated Adipocytes." J. Biol. Chem. 279: 35313-35319.

Rice, N.A., Nadeau, O.W., Yang, Q. and Carlson, G. M. (2002) “The Calmodulin-binding Domain of the Catalytic γ Subunit of Phosphorylase Kinase Interacts with Its Inhibitory α Subunit,” J. Biol. Chem. 277:14681-14687.

Nadeau, O.W. and Carlson, G.M. (2002). Chemical Crosslinking in Studying Protein-protein Interactions, in Protein-Protein Interactions, Chap. 6, Cold Spring Harbor Laboratory Press, New York, pp 75-91.

Nadeau, O.W., Domanski, P., Usacheva, A., Uddin, S., Platanias, L.C., Pitha, P., Raz, R., Levy, D., Majchrzak, B., Fish, E. and Colamonici, O.R. (1999) “The Proximal Tyrosines of the Cytoplasmic Domain of the β Chain of the Type I Interferon Receptor are Essential for Signal Transducer and Activator of Transcription (Stat) 2 Activation,” J. Biol. Chem. 274:4045-4052.

Nadeau, O.W., Traxler, K.W., Fee, L.R., Baldwin, B.B. and Carlson, G.M. (1999), “Activators of Phosphorylase Kinase Alter the Cross-linking of its Catalytic Subunit to the C-terminal 1/6th of its Regulatory α Subunit,” Biochemistry 38:2551-2559.

Carlson, G.M. and Nadeau, O.W. (1999) “Bifunctional Cross-linking Reagents,” in The Encyclopedia of Molecular Biology, Ed. T.E. Creighton, Wiley, New York, pp 279-281.

Carlson, G.M. and Nadeau, O.W. (1999) “Cross-linking,” in The Encyclopedia of Molecular Biology, Ed. T.E. Creighton, Wiley, New York, pp 582-584.

Carlson, G.M. and Nadeau, O.W. (1999) “Enzyme Immobilization and Conjugation,” in The Encyclopedia of Molecular Biology, Ed. T.E. Creighton, Wiley, New York, pp 829.

Carlson, G.M. and Nadeau, O.W. (1999) “Gluteraldehyde,” in The Encyclopedia of Molecular Biology, Ed. T.E. Creighton, Wiley, New York, pp 1019-1020.

Domanski, P., Nadeau, O.W., Platanias, L.C., Fish, E., Kellum, M., Pitha, P. and Colamonici, O.R. (1998), “Differential Use of the βL Subunit of the Type I Interferon Receptor Determines Signaling Specificity for IFNα2 and IFNβ,” J. Biol. Chem. 273:3144-3147.

Platanias, L.C., Domanski, P., Nadeau, O.W., Yi, T., Uddin, S., Fish, E., Neel, B.G. and Colamonici, O.R. (1998), “Identification of a Domain in the β Subunit of the Type I Interferon (IFN) Receptor that Exhibits a Negative Regulatory Effect in the Growth Inhibitory Action of Type I IFNs,” J. Biol. Chem. 273:5577-5581.

Ayers, N.A., Nadeau, O.W., Read, M.W., Ray, P. and Carlson, G.M. (1998), “Effector-sensitive Cross-linking of Phosphorylase-b Kinase by the Novel Cross-linker 4-phenyl-1,2,4-triazoline-3,5-dione,” Biochem. J. 331:137-141.

Nadeau, O.W., Traxler, K.W. and Carlson, G.M. (1998), “Zero-length Cross-linking of the β Subunit of Phosphorylase Kinase to the N-terminal Half of its Regulatory α Subunit,” Biochem. Biophys. Res. Commun. 251:637-641.

Nadeau, O.W., Sacks, D.B. and Carlson, G.M. (1997) “Differential Affinity Cross-Linking of Phosphorylase Kinase Conformers by the Geometric Isomers of Phenylenedimaleimide,” J. Biol. Chem. 272:26196-26201.

Nadeau, O.W., Sacks, D.B. and Carlson, G.M. (1997) “The Structural Effects of Endogenous and Exogenous Ca2+/Calmodulin on Phosphorylase Kinase,” J. Biol. Chem. 272:26202-26209.

Domanski, P., Fish, E., Nadeau, O.W., Witte, M., Platanias, L.C., Yan, H., Krolewski, J., Pitha, P. and Colamonici, O.R. (1997) “A Region of the β Subunit of the Interferon a Receptor Different from Box 1 Interacts With Jak1 and Is Sufficient To Activate the Jak-Stat Pathway and Induce an Antiviral State,” J. Biol. Chem. 272:26388-26393.

Nadeau, O.W., Falick, A. and Woodworth, R.C. (1996) Structural Evidence For An Anion-Directing Track in the Hen Ovotransferrin N-Lobe: Implications for Transferrin Synergistic Anion-Binding,” Biochemistry 35:14294-14303, 1996.

Nadeau, O.W. and Carlson, G.M. (1994) “Zero-length Conformation-dependent Cross-linking of Phosphorylase Kinase Subunits by Transglutaminase,” J. Biol. Chem. 269:29670-29676, 1994.