Research Interests:

A dynamic Ca2+ ion gradient formed across the sarcoplasmic reticulum (SR) membrane governs contraction and relaxation of the heart muscle cell. Muscle contraction is dependent on the movement of Ca2+ into the cytoplasm, whereas, relaxation requires the exclusion of Ca2+ ions from the cytoplasm. The finite balance of this gradient is regulated in part by an elaborate signaling cascade which has profound dependence on three proteins: cAMP-dependent protein kinase A (PKA), phospholamban (PLB), and sarcoendoplasmic reticulum Ca-ATPase (SERCA). My research is focused on solution NMR studies of the catalytic subunit of PKA. We are in pursuit of the full backbone assignment of PKA, the determination of its solution structure and dynamics, and finally the analysis of its interactions with PLB and PLB analogues.

Education:

Research Positions:

Teaching Positions:

Awards:

Publications:

Masterson, L.R., Bortone, N., Yu, T., Ha, K.N., Gaffarogullari, E., Nguyen, O., Veglia, G., Expression and Purification of Isotopically Labeled Peptide Inhibitors and Substrates of cAMP-dependent Protein Kinase A for NMR Analysis, Protein Expr. Purif., 2008, E-pub ahead of print.

Behling, L.A., Hartsel, S.C., Lewis, D.E., Dispirito, A.A., Choi, D.W., Masterson, L.R., Veglia, G., Gallagher, W.H. NMR, Mass Spectrometry and Chemical Evidence Reveal a Different Chemical Structure of Methanobactin that Contains Oxazolone Rings, J. Am. Chem. Soc., 2008, E-pub ahead of print.

Masterson, L.R., Tonelli, M., Markley, J.L., Veglia, G., Simultaneous Detection and Deconvolusion of Congested NMR Spectra Containing Three Isotopically Labeled Species, J. Am. Chem. Soc., 2008, 130, 25, 7818-19.

Masterson, L.R., Mascioni, A., Traaseth, N.J., Taylor, S.S., Veglia, G., Allosteric Cooperativitiy in Protein Kinase A, PNAS, 2008, 105, 2, 506-11.

Traaseth, N.J., Ha, K.N., Verardi, R., Shi, L., Buffy, J.J, Masterson, L.R., Veglia, G., Structural and Dynamic Basis of Phospholamban and Sarcolipin Inhibition of Ca(2+)-ATPase, Biochemistry, 2008, 47, 1, 3-13.

Tonelli, M., Masterson, L.R., Hallenga, K., Veglia, G., Markley, J.L., Carbonyl Carbon Label Selective (CCLS) (1)H-(15)N HSQC Experiment for Improved Detection of Backbone (13)C-(15)N Cross Peaks in Larger Proteins,J. Biomol. NMR, 2007, 39, 3, 177-85.

Masterson, L.R., Etienne, M.A., Porcelli, F., Barany, G., Hammer, R.P., Veglia, G., Non-stereogenic α-aminoisobutyryl-glycyl (Aib-Gly) Dipeptidyl Unit Nucleates Type I' β-turn in Linear Peptides in Aqueous Solution, Biopolymers, 2007, 88, 5, 746-53.

Gazal, S., Masterson, L.R., Barany, G., Facile Solid-phase Synthesis of C-terminal Peptide Aldehydes and Hydroxamates from a Common Backbone Amide-Linked (BAL) Intermediate, J. Pept. Res., 2005, 66, 6, 324-32.

Posters:

“Abberations in the Mechanism of Protein Kinase A Mediated Phosphorylation Caused by Phospholamban Mutants” (February 2008, 52nd Annual Biophysical Society Meeting- Long Beach, CA)

“Structural Basis for Dilated Cardiomyopathy” (January 2008, Keystone Symposia: Frontiers in Structural Biology- Steamboat, CO)

“Propagation of Internal Motions: The Catalytic Cycle of Protein Kinase A as Characterized by NMR” (March 2007, 51st Annual Biophysical Society Meeting- Baltimore, MD)

“Coupled Motions in Protein Kinase A as Characterized by NMR” (April 2006, Experimental Nuclear Magnetic Resonance Conference- Pacific Grove, CA; January 2007, Keystone Symposia: Frontiers of NMR in Molecular Biology- Snowbird, UT)

“Defining Allosteric Regulation in Protein Kinase A” (January 2006, Keystone Symposia: Frontiers in Structural Biology- Keystone, CO)

“Impact of a Single a,a -Disubstituted Amino Acid on b –Hairpin” (June 2005, 19th American Peptide Symposium- San Diego , CA )

“Unlocking the Structure and Flexibility in a Series of Peptides Using 1H NMR Spectroscopy” (March 2002, The 46 th Annual Biophysical Society Meeting- San Francisco , CA ; April 2002, National Conference of Undergraduate Research- Whitewater , WI )

“Studying the Structure and Flexibility of a Short 310 –helical Peptide using 1H NMR Spectroscopy” (April 2001 and 2002, UW-L Celebration of Undergraduate Research and Creativity- La Crosse, WI)

Research Support:

American Heart Association Predocotoral Fellowship, “Molecular and Structural Basis of Dilated Cardiomyopathy: An Atomic Investigation of Key Protein-Protein Interactions” (7/1/06-6/31/07)

Goals: Solid-phase peptide synthesis of phospholamban (PLN) analogues and protein kinase substrates. Solution NMR assignment of C-subunit PKA, characterization of binding standard ligands, and optimization of kinetic measurements in the presence of lipids. Characterization of the interactions with peptide models corresponding to PLN and R9C-PLN.
Role: Principle Investigator

NIH-Chemical Biology Interface Training Grant, “Origins of Dilated Cardiomyopathy: An Atom-by Atom Investigation of Dynamic Processes in Key Enzymes” (7/2005-7/2006)

Goals: Solid-phase peptide synthesis of phospholamban analogues and protein kinase substrates. Optimization for the expression and purification of C-subunit PKA NMR samples.
Role: Trainee

Goals: Solution state peptide synthesis of short 310-helical peptides and use of NMR to characterize the thermodynamic stability of their secondary structure.
Role: Researcher