Research and Publications

David L. Daleke

Associate Professor, Biochemistry and Molecular Biology
Associate Dean, University Graduate School

B.S., Stanford University, 1979; Ph.D. Stanford University, 1986.
American Cancer Society Postdoctoral Fellow, University of California at San Francisco, 1986-88; Fellow, Indiana Molecular Biology Institute, 1988; Established Investigator, American Heart Association, 1994-1999; Indiana University School of Medicine Eminent Scholar, 1999.

Medical Sciences
Myers Hall 200
915 E. 3rd Street
Indiana University
Bloomington, Indiana 47405

Research Interests

The generation and maintenance of transmembrane phospholipid asymmetry are essential for the function of biological membranes, yet the mechanism underlying these fundamental processes remains unclear. Our research effort is designed to understand how phospholipids are assembled in biological membranes and how the resulting phospholipid asymmetry is maintained. Phospholipid transporters, or "flippases," such as the recently discovered aminophospholipid translocator, represent an interesting new class of proteins that may play a key role in the assembly and organization of phospholipids in biological membranes. These enzymes require energy in the form of ATP, have strict phospholipid structural requirements, and are unique in their ability to transport lipids across membranes. Our goal is to elucidate the structure, function, and biological significance of these proteins.

Our studies employ a variety of biochemical, biophysical, and spectroscopic methods, including protein chemistry, radiolabel, and fluorescent techniques. Part of our work is directed at purifying the aminophospholipid flippase from human erythrocyte membranes. We have purified an ATPase that bears physical characteristics consistent with its involvement in aminophospholipid transport. Our enzymological studies have shown that this enzyme is specifically simulated by phosphatidylserine, the primary substrate of the aminophospholipid flippase. Once this transporter is reconstituted into model membranes, further biophysical studies of lipid-protein interactions and molecular mechanisms of phospholipid transport will be performed.

Concurrently, we are investigating the role of blood cell membrane structure in cardiovascular disease. Specifically, we are studying the loss of transmembrane phospholipid asymmetry observed in diabetic red blood cells to determine the relationship between the vascular complications associated with diabetes and membrane structural perturbations. Our studies indicate that hyperglycemic treatment of non-diabetic cells duplicates this loss of asymmetry by increasing passive lipid flip-flop, without affecting aminophospholipid flippase activity. Antioxidants suppress this loss of asymmetry, implicating a role for glucose-mediated lipid oxidation. Ongoing studies are designed to determine the mechanism by which lipid oxidation induces membrane lipid scrambling, including studies with animal models of diabetes and human diabetics. In related work, we are studying phospholipid transport in normal and diabetic blood platelets to understand the role of oxidative inhibition of the flippase in aminophospholipid externalization, a process required for normal blood clotting. These studies may lead to the development of new strategies for the treatment and prevention of heart disease.

Publications

Jennifer A. Meyer, Wasanthi Subasinghe, Anders A. F. Sima, Zachary Keltner, Gavin E. Reid, David Dalekeand Dana M. Spence, “Zinc-activated C-peptide Resistance to the Type 2 Diabetic Erythrocyte is Associated with Hyperglycemia-induced Phosphatidylserine Externalization and Reversed by Metformin,” Molecular BioSystems (2009).

David L. Daleke, “Regulation of Phospholipid Asymmetry in the Erythrocyte Membrane,” Current Opinion in Hematology 15 (2008) 191-195. PMID: 18391783.

Smriti and David L. Daleke, “ATP-dependent Transport of Phosphatidylserine Analogs in Human Erythrocytes,” Biochemistry, 46 (2007) 2249-2259. PMID: 17269657.

David L. Daleke, “Phospholipid Flippases”, Journal of Biological Chemistry 282 (2007) 821-825 (invited review). PMID: 17130120.

Jill Paterson, Kathleen Renkema, Maragaret Halleck, Robert Schlegel, Patrick Williamson and David L. Daleke, Lipid Specific Activation of the Murine P4-ATPase Atp8a1 (ATPase II),” Biochemistry 45 (2006) 5367-5376. PMID: 17130120.

Dana M. Niedowicz and David L. Daleke, “The Role of Oxidative Stress in Diabetic Complications,” Cell Biochemistry and Biophysics 43 (2005) 289-330. PMID: 16049352.

Peter R. Hoffmann, Jennifer A. Kench, Andrea Vondracek, Ellen Kruk, David L. Daleke, Michael Jordan, Philippa Marrack, Peter M. Henson, and Valerie A. Fadok, “Interaction between Phosphatidylserine and the Phosphatidylserine Receptor Inhibits Immune Responses In Vivo.” J Immunol 174 (2005) 1393-1404. PMID: 15661897.

Jill V. Lyles, Kathleen Cornely-Moss, Christine M. Smith and David L. Daleke, “Purification of the Human Erythrocyte PS-Stimulated Mg²⁺sup-ATPase: A Putative Flippase,” Methods in Molecular Biology 228 (2003) 257-269. PMID: 12824559.

David L. Daleke, “Regulation of Transbilayer Plasma Membrane Phospholipid Asymmetry,” Journal of Lipid Research 44 (2003) 233-242. PMID: 12576505.

Mou Banerjee, Daryl C. Drummond, Arvind Srivastava, David L. Daleke, and Barry R. Lentz, “Specificity of Soluble Phospholipid Binding Sites on Human Factor Xa,” Biochemistry 41 (2002) 7751-7762. PMID: 12056907.

Xin Zhai, Arvind Srivastava, Daryl C. Drummond, David Daleke, and Barry R. Lentz, “Phosphatidylserine Binding Alters the Conformation and Specifically Enhances the Cofactor Activity of Bovine Factor Va,” Biochemistry 41 (2002) 5675-5684. PMID: 11969429.

Peter R. Hoffmann, Aimee M. deCathelineau, Carol Anne Ogden, Yann Leverrier, Donna L. Bratton, David L. Daleke, Anne J. Ridley, Valerie A. Fadok, and Peter M. Henson, “Phosphatidylserine (PS) Induces PS receptor–mediated Macropinocytosis and Promotes Clearance of Apoptotic Cells,” Journal of Cell Biology 155 (2001) 649-659. PMCID: PMC2198875. PMID: 11706053.

Valerie A. Fadok, Aimee de Cathelineau, David L. Daleke, Peter M. Henson and Donna L. Bratton, “Loss of Phospholipid Asymmetry and Surface Exposure of Phosphatidylserine is Required for Phagocytosis of Apoptotic Cells by Macrophages and Fibroblasts,” Journal of Biological Chemistry 276 (2001) 1071-1077. PMID: 10986279.

Umakant J. Dumaswala, Michael J. Wilson, Y. L. Wu, J. Wykle, L. Zhuo, L. M. Douglass, and David L. Daleke, “Glutathione Loading Prevents Free Radical Injury in Red Blood Cells After Storage,” Free Radical Research 33 (2000) 517-529. PMID: 11200085.

David L. Daleke and Jill V. Lyles, “Identification and Purification of Phospholipid Flippases,” Biochimica et Biophysica Acta 1486 (2000) 108-127. PMID: 10856717.

Kayoko Uchida, Kazuo Emoto, David L. Daleke, Keizo Inoue, and Masato Umeda, " Induction of Apoptosis by Phosphatidylserine," Journal of Biochemistry 123 (1998) 1073-1078. PMID: 9603995.

Joanne E. Johnson, Michael L. Zimmerman, David L. Daleke, and Alexandra C. Newton, “Lipid Structure and Not Membrane Structure Is the Major Determinant in the Regulation of Protein Kinase C by Phosphatidylserine,” Biochemistry 37 (1998) 12020-12025. PMID: 9724512.

Umakant J. Dumaswala, Michael Wilson, Thomas José, and David L. Daleke, “Effect of a Glycerol-Containing Hypotonic Medium on Phospholipid Asymmetry and Aminophospholipid Transport During Storage," Biochimica et Biophysica Acta 1330 (1997) 265-273. PMID: 9408180.

Umakant J. Dumaswala, Michael J. Wilson, Thomas José, and David L. Daleke, "Glutamine- and Phosphate-Containing Hypotonic Storage Media Better Maintain Erythrocyte Membrane Physical Properties," Blood 88 (1996) 697-704. PMID: 8695818.

David L. Daleke, Jill V. Lyles, Edward Nemergut and Michael Zimmerman, "Purification and Substrate Specificity of the Human Erythrocyte Aminophospholipid Transporter," in Trafficking of Intracellular Membranes: From Molecular Sorting to Membrane Fusion, Maria Pedroso de Lima, Dick Hoekstra and Nejat Düzgünes, Eds., Springer-Verlag, New York, 1995, pp 49-59.

Daryl Drummond and David L. Daleke, "Synthesis and Characterization of pH-Dependent "Caged" Aminophospholipids," Chemistry and Physics of Lipids 75 (1995) 27-41. PMID: 7697781.

David L. Daleke, Wray H. Huestis, and Alexandra C. Newton, "Protein Kinase C as a Measure of Transbilayer Phosphatidylserine Asymmetry," Analytical Biochemistry 217 (1994) 33-40. PMID: 8203737.

Michael Zimmerman and David L. Daleke, "Regulation of a Candidate Phosphatidylserine-Transporting ATPase by Lipid," Biochemistry 32 (1993) 12257-12263. PMID: 8218304.

Michael J. Wilson, Kay Richter-Lowney, and David L. Daleke, "Hyperglycemia Induces a Loss of Phospholipid Asymmetry in Human Erythrocytes," Biochemistry 32 (1993) 11302-11310. PMID: 8218195.

Hoai-Thu N. Truong, David L. Daleke, and Wray H. Huestis, "Dithiothreitol Stimulates the Activity of the Plasma Membrane Aminophospholipid Translocator," Biochimica et Biophysica Acta 1150 (1993) 57-62. PMID: 8334138.

Hoai-Thu N. Truong, David L. Daleke, and Wray H. Huestis, "Human Erythrocyte Shape Regulation: Interaction of Metabolic and Redox Status," Biochimica et Biophysica Acta 1150 (1993) 51-56. PMID: 8334137.

David L. Daleke, Kathleen Cornely-Moss, Jill Lyles, Christine M. Smith, and Michael Zimmerman "Identification and Characterization of a Candidate Phosphatidylserine-Transporting ATPase," Annals of the New York Academy of Sciences 671 (1992) 468-470. PMID: 1288343.

Paul Meers, David L. Daleke, Keelung Hong, and Demetrios Papahadjopoulos, "Interactions of Annexins with Membrane Phospholipids," Biochemistry 30 (1991) 2903-2908. PMID: 1826088.

Robert E. Smith and David L. Daleke "Phosphatidylserine Transport in Rhnull Erythrocytes," Blood 76 (1990) 1021-1027. PMID: 2118395.

David L. Daleke, Keelung Hong, and Demetrios Papahadjopoulos, "Endocytosis of Liposomes by Macrophages: Binding, Acidification and Leakage of Liposomes Monitored by a New Fluorescence Assay," Biochimica et Biophysica Acta 1024 (1990) 352-366. PMID: 2162207.

Kristen M. Hege, David L. Daleke, Thomas A. Waldmann, and Katherine K. Matthay, "Comparison of Anti-Tac and Anti-Transferrin Receptor-Conjugated Liposomes for Specific Drug Delivery to Adult T Cell Leukemia," Blood 74 (1989) 2043-2052. PMID: 2679914.

David L. Daleke and Wray H. Huestis, "Erythrocyte Morphology Reflects the Transbilayer Distribution of Incorporated Phospholipids," Journal of Cell Biology 108 (1989) 1375-1385. PMID: 2925790.

David L. Daleke and Wray H. Huestis, "Incorporation and Translocation of Aminophospholipids in Human Erythrocytes," Biochemistry 24 (1985) 5406-5416. PMID: 4074704.

Mark J. Nelson, David L. Daleke, and Wray H. Huestis, "Calmodulin-dependent Spectrin Kinase Activity in Resealed Human Erythrocyte Ghosts," Biochimica et Biophysica Acta, 686 (1982) 182-188. PMID: 6805511.

revised August 2009