Signal Transduction in Blood Cells, Normal and Leukemic

• News release: "Tribute Given to Dr. Gomez-Cambronero," 2005.
• News release: "WSU Researcher at the Forefront of a New Field," 2004.
• Vital Signs article: "Researchers Seek to Unravel a Central Paradigm in Cell Biology," 2004.

Research interests:

Molecular cell signaling in human neutrophils as elicited by hematopoietic growth factors; cell migration (chemotaxis); pathological connections with inflammation-mediated tissue injury and with leukemia; new methodologies for the study of protein-protein interaction and enzyme inhibitor design.

Research lines:

1. Neutrophil/Endothelium/Pathogen injury & atherogenesis
2. Neutrophil PLD, oxygen radicals, & ischemia/reperfusion
3. Signal transduction pathways in leukemic cells

Students' achievements

Madhu Mahankali

• Received the Biomedical Sciences Ph.D. program's Graduate Student Excellence Award for the 2009-10 academic year.
• Received won the Best oral scientific presntation at Biomedical Science Retreat, 2011

Amy Price

• Best Poster Presentation (Science Apprenticeship Program)
• The Mead Westvaco College Tuition Award

Isabel Campos

• Best Poster presentation (STREAMS Program, School of Medicine)
• Travel Award, Experimental Biology Meeting, Washington, D.C.

Mauricio Di Fulvio, Ph.D.

• Travel Award, 36th National Meeting of the Society for Leukocyte Biology, Philadelphia, PA
• Best oral presentation at the 10th Annual WSU Molecular Biology Retreat

Research line 1:
Neutrophil/Endothelium/Pathogen injury and atherogenesis

For some time, researchers have been awestruck with the fact that nearly 50 percent of the people having atherosclerosis and heart attacks had normal levels of cholesterol and oxidized LDL and followed healthy lifestyles. In the last few years, investigators seem to have identified a new "culprit" for the remaining 50 percent: Inflammation. The connection of the immune system's powerful inflammatory response with cardiopathies are now being increasingly recognized. Further, there is considerable evidence that pathogens from the oral flora can cause damage in coronary arteries (the Red Cross will not accept a blood donation from anyone who has had a dental cleaning in the previous 24 hours).

Based on this, we have proposed that bacterial infection of endothelial cells lining the coronary artery's wall will create a focus of infection. This will attract neutrophils from the bloodstream. The neutrophils' mission is to seek and destroy invading pathogens. Initially, neutrophils will attempt to eliminate the invading pathogen in the artery by the release of powerful proteases and reactive oxygen species. But in doing so, these very substances will inflict collateral damage on the endothelial and fibroblast walls. The problem is that in predisposed individuals, this process might become chronic, particularly with the recruitment and the arrival of more leukocytes and the overall amplification by numerous inflammatory signals (cytokines). If the process does not resolve appropriately, this "battlefield" will leave a "scar" of damaged tissue in the lining of the artery which will serve as the nidus for a nascent atherosclerotic plaque.

Pathogens, inflammation and atherogenesis hypothesis

We have proposed to develop and establish a "Neutrophil-Endothelial Cell-Pathogen" (N EC-P) model. This represents a completelynovel experimental model of an artery wall for the purpose of identifying chemoattractant molecules released by infected EC and assessing the enzymatic damage produced by activated neutrophils. This will comprise:

[a]Bacterially-infected endothelial cells (ECs) with S. sanguisand S. epidermidis, [b]ECs' release of cytokines IL-8 and GM-CSF, [c]Neutrophil infiltration to the site of infection, [d]Destruction of bacteria by myeloperoxidase (MPO) and other lytic enzymes, and [e] Collateral damage inflicted upon ECs and other cells (namely, fibroblasts and smooth muscle cells).

This model will provide new molecular targets for the interaction of infected vascular endothelium and cells of the innate immune system. These in turn could be used to design treatments to prevent neutrophil mediated tissue injury that occurs in atherosclerosis.

Techniques for this research

In vitromodels of atherogenesis; cell co-cultures; cDNA array techniques to pinpoint the genes involved in cytoskeletal-related signaling pathways; co-immunoprecipiation and in vitrokinase assays; forced expression of genes and their effect on parallel signaling pathways; transfection of dominant negative cDNA, antisense oligonucleotides and renatured proteins and assessment of pathophysiological effects.

Research line 2:
Neutrophil phospholipase D (PLD), oxygen radicals, and ischemia/reperfusion injury

We are seeking to understand the molecular mechanism underlying the production of lethal oxygen radicals duringischemia and reperfusion injury. Our studies focus on the enzyme phospholipase D (PLD) that is central to the production of phosphatidic acid (PA), a putative second messenger involved in the release of superoxide anions by neutrophils' respiratory burst (NADPH oxidase system). There are two fronts of attack in this line:

[a] To purify and clone neutrophil PLD, which we propose to be a distinctive form from the described mammalian PLD1 and PLD2 isoforms.Our first aim is to purify the enzyme by fast protein liquid chromatography (FPLC) and by molecular biology techniques, using the N-terminal 15-20 amino acids of the sequenced purified protein as well as the existing mammalian PLD cDNA sequences (PLD1 and PLD2). Our second aim is to determine the mechanism of granulocyte PLD regulation, particularly the phosphorylating kinase, the site(s) of phosphorylation and the role of small GTP-binding proteins, using immunoprecipitation with antibodies against epitope-tagged proteins and in vitroenzymatic assays.

Our working hypothesis is that a novel isoform of PLDin human neutrophils is regulated through tyrosine phosphorylation, which allows the enzyme to become active and synthesize phosphatidic acid (PA), that in turn triggers the release of toxic oxygen radicals. These are responsible for the exacerbation of tissue damage and necrosis after a heart attack.

[b] To find a selective inhibitor of PLD,intended to ameliorate the devastating effectsof the ischemia/reperfusion pathological condition. This will be done in the following sequential steps:

Techniques for this research

Organic synthesis by combinatorial chemistry methods; fluorography; amino acid and tryptic peptide analyses; peptide arrays; generation of protein single-point mutants; three-dimensional structure modeling and other proteomics methodologies on protein-protein interaction and enzyme inhibitor design.

Funding

This research line is funded by the American Heart Association (National Program).

Research line 3:
Signal transduction pathways in leukemic cells

We are seeking to understand the signal transduction mechanisms that are initiated by the binding of the granulocyte-macrophage colony stimulating factor (GM-CSF) to human leukocytes (leukemic blasts and mature neutrophils).

This hematopoietic growth factor is currently used to restore the normal white blood cells circulating levels in patients undergoing chemotherapyand during bone marrow transplantation. GM-CSF stimulates major signaling pathways such as mitogen-activated protein kinases (MAPK) p42ERK2 and p44ERK1, which become dually-phosphorylated on tyrosine and serine/threonine; and the ribosomal S6 kinases p90rsk and p70S6K.

Positive feedback of MAPK on p70s6k pathway leading to heightened chemotaxis

Using proleukemic cell lines of the myelocytic lineage as the biological source, we are currently studying:

[a]the point at which MAP and S6 kinases are turned on/off during the late stages of white blood cell maturation; and

[b]the physiological role of MAPK upregulation in mature cells. To this, our data aim at a molecular cross-talkbetween MAPK and p70S6K signaling pathways.

We propose that upon commitment to hematopoietic differentiation, the normal transduction pathways switch from cell-dividing mechanisms to a way that allows the cell to mount a full response (e.g., chemotaxis and phagocytosis) against invading pathogen during infection.

Techniques for this research

Leukemic cell culture; immunochemistry (phosphotyrosine analyses); RT-PCR; cytopreprarations; flow cytometry; immunofluorescence microscopy; enzyme assays (radiometric, photometric and in-gel); phagocyte functional assays.

Funding

This research line is funded by Wright State University Boonshoft School of Medicine and the National Institutes of Health (NIH-NHLBI).

Dr. Julian Gomez-Cambronero

Dr. Julian Gomez-Cambronero received his Ph.D. (cum laude) in Biochemistry/Immunology at the Complutense University (Madrid, Spain) in 1986 under Drs. M. Sanchez Crespo and J. M. Mato's direction. He then moved to the United States and completed his training in Cell Biology as a postdoctoral fellow in Dr. R. I. Sha'afi's laboratory at the Department of Physiology of the University of Connecticut Health Center, and with the late Dr. E. L. Becker. From 1991 to 1995 he was part of the faculty at the University of Connecticut, first as an Instructor and then as a Research Assistant Professor. In 1995 he moved to the Department of Physiology and Biophysics at Wright State University, chaired by Dr. P.K. Lauf, to accept a tenure-track Assistant Professorship position, and was promoted to Associate Professor in 2000 and to full Professor in 2004. He is married to Teresa Madrid and has two children, David and Julia.

Member of the graduate program

As part of the graduate faculty and as a dissertation-qualified professor, he participates in the following areas of concentration of the Biomedical Sciences (BMS) Doctoral (Ph.D.) Program: Cell Biology and Physiology, Immunology, and Biochemistry and Molecular Biology. He also participates in the Master of Science (MS) Program in Physiology and Biophysicsand the Immunology MS Program. His formal lectures include an "Introduction to Hematology" for first year medical students in the team-taught Molecular, Cellular and Tissue Biology course; "Molecular Basis of Leukemia and Lymphoma" second year medical students; and "Signal Transduction/Molecular Endocrinology and Blood Physiology" for the BMS Ph.D. Program. He is course director for "Blood/Hematology" for the School of Medicine.

Honors and awards

Dr. Cambronero is a full member of the American Physiological Society, the American Society for Biochemistry and Molecular Biology, the American Society of Hematology, and the Society for Leukocyte Biology. He received an American Heart Association (AHA) Postdoctoral Fellowship in 1987; was part of the Leukemia Society of America (LSA) Northern Connecticut Chapter Telecast in 1990; was National Finalist for the Young Investigator Award of the Society for Leukocyte Biology (SLB) in 1992; received the New Investigator Research Award from the Donaghue Medical Research Foundation in 1992; was elected as Expert Scientist by the Farmington, Connecticut, Public School Board to evaluate the science curriculum in 1994; was the subject of a biographical record in the American Men & Women of Science directory in 1995; received the Frontiers in Physiology National Research Award from the American Physiological Society (APS) in 1996; was recognized with the STARS Scholar Distinguished Service Award by The Ohio University in 1998; received the Professional Achievement "Sembrador" Award from The City of Manzanares (CR), Spain, in 2004; and received the Outstanding Achievement in Medical Education and Research Award from the Academy of Medicine of Dayton in 2007.

Funding and scholarly publications

Dr. Cambronero has received funding from the National Institutes of Health (NIH NHLBI); the American Cancer Society (ACS); the American Heart Association (National Program); the American Physiological Society; the Ohio Board of Regents; the University of Connecticut; and Wright State University.

He has been invited to the Erythrocyte and Leukocyte Biology (ELB), Cell Development and Function (CDF-2), and Innate Immunity and Inflammation (III) Study Sections for the NIH; and to the Molecular Signaling, Basic Cell and Molecular Biology Review Panel for the American Heart Association
(AHA) Research Consortium.

He has published over 60 original research papers in peer-review journals, five book chapters, and has presented over 50 research posters and seminar talks both nationally and internationally. He has also published science-fiction short stories and layman science-related articles in Spanish literary magazines.

Oganizing a grant-writing workshop

Dr. Cambronero is the chair of the Professional Development Committee of the Society of Leukocyte Biology (SLB) whose mission is to provide educational opportunities for junior scientists and immunologists. He developed and organized the Annual Workshop on "How to Write Your First Research Grant" with advice for grad students and junior professors on grantsmanship for NIH grants, which was held for the first time in Kansas City, Mo., in September 2011.

Making science available to the general public

Dr. Cambronero came up with the idea and designed el Paseo del Sistema Solar (A walk through the Solar System) in the City of Manzanares (Castilla-La Mancha, Spain). It is a scale model of the Solar System, where representations of the Sun and the planets have been positioned along a path throughout a public park, according to a scale that reduces the millions of miles in the cosmos down to yards in the park. The actual representations of the Sun and planets are spheres constructed of fiberglass, set within rectangular prisms or "monoliths" of oxidized steel, measuring about six feet tall each. The objective of the "Paseo del Sistema Solar" is for the visitor to gain an understanding and appreciation of the workings of the Solar System in a "hands-on," "walk-through" model. Dr. Cambronero wanted to make sure that science is explained and made available to the general public. He hopes this project particularly reaches the younger generations, encouraging them to become interested in the wonders of science. Visit http://www.paseodelsistemasolar.manzanares.es for more information.

PUBLICATIONS

Representative of Doctorate Period:

Gomez-Cambronero, J., Mato, J.M., Vivanco, F. and Sanchez-Crespo, M. (1987) Phosphorylation of partially purified 1-0-alkyl-2-lyso-glycero-3-phosphocholine: acetyl-CoA acetyltransferase from rat spleen. Biochem. J.245, 893-898. PDF

Representative of Postdoctoral Period:

Gomez-Cambronero, J., Huang, C-K., Gomez-Cambronero, T.M., Waterman, W.H., Becker, E.L. and Sha'afi, R.I. (1992) Granulocyte-macrophage colony-stimulating factor-induced protein tyrosine phosphorylation of MAP Kinase in human neutrophils. Proc. Natl. Acad. Sci.USA. 89, 7551-7555. PDF

As Principal Investigator:

Johnson, G.M. and Gomez-Cambronero, J. (1995) Priming of tyrosine phosphorylation in GM-CSF-stimulated adherent neutrophils. J. Leukocyte Biol.57, 692-698. Abstract

Gomez-Cambronero, J. (1995) Immunoprecipitation of a phospholipase D activity with anti-phosphotyrosine antibodies. J. Interferon Cytok. Res.15, 877-885. Abstract

Joseph, D., Paul, C.C., Baumann, M.A. and Gomez-Cambronero, J. (1996) S6 kinase p90rsk in GM-CSF-stimulated proliferative & mature hemopoietic cells. J. Biol. Chem.271, 13088-93. PDF

Gomez-Cambronero, J. and Veatch, C. (1996) Emerging paradigms in granulocyte macrophage colony-stimulating factor signaling. Life Sci.59, 2099-2111. Abstract

Gomez-Cambronero, J. and Keire, P. (1998) Phospholipase D: a novel major player in signal transduction. Cell Signal. 10, 387-397.

Hayes, T.S., Billington, C.J., Robinson, K.A., Sampt, E.R., Fernandez, G.A. and Gomez Cambronero, J. (1999) Binding of granulocyte-macrophage colony-stimulating factor to adherent neutrophils activates Phospholipase D. Cell Signal.11, 195-204.

Gomez-Cambronero, J. (1999) MAP kinase is activated in EGF-stimulated interphase but not in mitotic HeLa cells. FEBS Lett.443, 126-130. PDF

Grishin, A., Sinha, S., Roginskaya, V., Boyer, M.J., Gomez-Cambronero, J., Zuo, S., Kurosaki, T., Romero, G. and Corey S.J. (2000) Involvement of Shc and Cbl-PI 3 kinase in Lyn-dependent proliferative signaling pathways for G-CSF. Oncogene19, 97 105. Abstract

Andrews, B., Bond, K., Lehman, J.A., Horn, J.M., Dugan, A. and Gomez-Cambronero, J. (2000) Direct inhibition of in vitro PLD activity by 4-(2-aminoethyl) benzenesulfonyl fluoride. Biochem. Biophys. Res. Com.273, 302-311.

Paul, C.C., Aly, E., Lehman, J.A., Page, S.M., Gomez-Cambronero, J., Ackerman, S.J. and Baumann, M.A. (2000). Human cell line that differentiates to all myeloid lineages and expresses neutrophil secondary granule genes. Exp. Hematol.28, 1373-1380.

Lehman, J.A., Paul, C.C., Baumann, M.A. and Gomez-Cambronero, J. (2001) MAP kinase upregulation after hematopoietic differentiation: role of chemotaxis. Am. J. Physiol. Cell Physiol.280, 183-191. PDF

Horn, J.M., Lehman, J.A., Alter, G., Horwitz, J. and Gomez-Cambronero, J. (2001) Presence of a phospholipase D (PLD) distinct from PLD1 or PLD2 in human neutrophils: immuno-biochemical characterization and initial purification. Biochim. Biophys. Acta1530, 97-110.

Blum, J.J., Lehman, J.A., Horn, J.M. and Gomez-Cambronero, J. (2001) Phospholipase D (PLD) is present in L. donovaniand its activity increases in response to acute osmotic stress. J. Eukaryotic Microbiol.48, 102-110. Abstract

Sampt, E.R., Fernandez, G.A., Lehman, J.A., Corey, S.J., Huang, C.-K. and Gomez Cambronero, J. (2001) A systematic approach to the complete study of a signaling molecule: ribosomal p90rsk as an example.J. Biochem. Biophys. Meth.48, 219-237.

Gomez-Cambronero, J. (2001) The oxygen dissociation curve of hemoglobin: bridging the gap between Biochemistry and Physiology. J. Chem. Ed.78, 757-759. Abstract

Baumann, M.A., Paul, C.C., Lemley-Gillespie, S., Oyster, M. and Gomez-Cambronero, J. (2001) Modulation of MEK activity during G-CSF signaling alters proliferative vs.differentaitive balancing.Am. J. Hematol.68, 99-105. Abstract

Lehman, J.A. and Gomez-Cambronero, J. (2002) Molecular crosstalk between p70S6K and MAPK cell signaling pathways. Biochem. Biophys. Res. Com.293, 463-469.

Gomez-Cambronero, J., Horwitz, J. and Sha'afi, R.I. (2003) Measurements of Phospholipases A2, C and D (PLA2, PLC and PLD): in vitro microassays, analysis of enzyme isoforms and intact cells assays. Methods Mol. Biol.218, 155-1576. Abstract

Lehman, J.A., Calvo, V. and Gomez-Cambronero, J. (2003) Mechanism of ribosomal p70S6 kinase activation by GM-CSF in neutrophils: Cooperation of a MEK-related, T421/S424-kinase and a rapamycin-sensitive, mTOR-related, T389-kinase. J. Biol. Chem.278, 28130-28138. PDF

Gomez-Cambronero, J. (2003) Rapamycin inhibits GM-CSF-induced neutrophil migration. FEBS Let.550, 94-100.

Gomez-Cambronero, J., Horn, J., Paul, C.C. and Baumann, M.A. (2003) GM-CSF is a chemoattractant cytokine for neutrophils: Involvement of the ribosomal p70S6K signaling pathway. J. Immunol.171, 6846-6855. Abstract

Gomez-Cambronero, J., Frye, T. and Baumann, M.A. (2004) Ribosomal p70S6K basal activity increases upon induction of differentiation of myelomonocytic leukemic cell lines HL60, AML14 and MPD. Leukemia Res.28, 755-62.

Baumann M, Frye T, Naqvi T and Gomez-Cambronero J. (2005) Normal neutrophil maturation is associated with selective loss of MAP kinase activation by G-CSF. Leukemia Res.29, 73-8.

Di Fulvio, M. and Gomez-Cambronero, J. (2005) Phospholipase D gene expression in human neutrophils and HL-60 differentiation. J. Leukocyte Biol.77, 999-1007. Abstract

Horn, J., Miller, M. Lopez, I. and Gomez-Cambronero, J. (2005) The uncovering of a novel regulatory mechanism for PLD2: Formation of a ternary complex with Protein Tyrosine Phosphatase PTP1b and Growth Factor Receptor-Bound Protein GRB2. Biochem. Biophys. Res. Com.332, 58-67. Abstract

Di Fulvio, M., Lehman, N., Xiaohong Lin, X, Lopez, I. and Gomez-Cambronero, J. (2006) The elucidation of novel SH2 binding sites on PLD2. Oncogene 25, 3032-3040. Abstract.

Lehman N, Di Fulvio M, McCray N, Campos I, Tabatabaian F, Gomez-Cambronero J. (2006) Phagocyte cell migration is mediated by phospholipases PLD1 and PLD2. Blood108, 3564-72. Abstract.

Lehman N, Ledford B, Di Fulvio M, Frondorf K, McPhail LC, Gomez-Cambronero J. (2007) Phospholipase D2-derived phosphatidic acid binds to and activates ribosomal p70 S6 kinase independently of mTOR. FASEB J 21, 1075-1087. Abstract.

Di Fulvio M, Frondorf K, Henkels KM, Lehman N, Gomez-Cambronero J. (2007) The Grb2/PLD2 interaction is essential for lipase activity, intracellular localization and signaling in response to EGF. J Mol Biol. 367, 814-824. Abstract.

Gomez-Cambronero J, Di Fulvio M, Knapek K. (2007) Understanding phospholipase D (PLD) using leukocytes: PLD involvement in cell adhesion and chemotaxis. J Leukoc Biol. 82, 272- 281. Abstract.

Di Fulvio M, Henkels KM, Gomez-Cambronero J. (2007) Short-hairpin RNA-mediated stable silencing of Grb2 impairs cell growth and DNA synthesis. Biochem Biophys Res Commun. 357, 737-742. Abstract.

Fulvio MD, Frondorf K, Gomez-Cambronero J. (2008) Mutation of Y(179) on phospholipase D2 (PLD2) upregulates DNA synthesis in a PI3K-and Akt-dependent manner. Cell Signal. 20, 176-185. Abstract.

Lehman, N., Ledford, B., Di Fulvio, M., Frondorf, K., McPhail, L.C. and Gomez-Cambronero, J. (2007) Phospholipase D2-derived phosphatidic acid binds to and activates ribosomal p70S6 Kinase independently of mTOR. FASEB J. 21, 1075-1087.

Di Fulvio, M., Frondorf, K., Henkels, K.M., Lehman, N. and Gomez-Cambronero, J. (2007) The Grb2/PLD2 interaction is essential for lipase activity, intracellular localization and signaling in response to EGF. J. Mol. Biol. 367, 814-824.

Di Fulvio, M., Henkels, K. and Gomez-Cambronero, J. (2007) Short-hairpin RNA-mediated stable silencing of Grb2 impairs cell growth and DNA synthesis. Biochem. Biophys. Res. Commun. 357, 737-742.

Gomez-Cambronero, J. Di Fulvio, M. and Knapek, K. (2007) Understanding Phospholipase D (PLD) using leukocytes: PLD involvement in cell chemotaxis and adhesion. J. Leukocyte Biol. 82, 272-281.

Di Fulvio, M., Frondorf, K. and Gomez-Cambronero, J. (2008) Mutation of Y179 on phospholipase D2 (PLD2) upregulates DNA synthesis in a PI3Kand Akt-dependent manner. Cell Signaling 20, 176-185. Information from this paper has been curated (2008) into PhosphoSite.

Haines, E., Minoo, P., Feng, Z., Resalatpanah, N., Nie, X.M., Campiglio, M., Alvarez, L., Cocolakis, E., Ridha, M., Di Fulvio, M., Gomez-Cambronero, J., Lebrun, J.J. and Ali, S. (2009) Tyrosine Phosphorylation of Grb2: Role in Prolactin/Epidermal Growth Factor Crosstalk in Mammary Epithelial Cell Growth and Differentiation. Mol. Cell. Biol. 29, 2505-2520.

Henkels, K.M., Short S., Peng, H.-J., Di Fulvio, M. and Gomez-Cambronero, J. (2009) PLD2 has both enzymatic and cell proliferation-inducing capabilities, that are differentially regulated by phosphorylation and dephosphorylation. Biochem. Biophys. Res. Commun. 389, 224-228.

Henkels, K.M., Peng, H.-J., Frondorf, K. and Gomez-Cambronero, J. (2010) A comprehensive model that explains the regulation of Phospholipase D2 (PLD2) activity by phosphorylation-dephosphorylation. Mol. Cell Biol. 30, 2251-2263.

Knapek K, Frondorf K, Post J, Short S, Cox D, Gomez-Cambronero J. (2010) The molecular basis of phospholipase D2-induced chemotaxis: elucidation of differential pathways in macrophages and fibroblasts. Mol. Cell. Biol. 30, 4492-506.

Gomez-Cambronero J. (2010) New concepts in phospholipase D signaling in inflammation and cancer. ScientificWorldJournal. 10, 1356-69

Tabatabaian F, Dougherty K, Di Fulvio M, Gomez-Cambronero J. (2010) Mammalian target of rapamycin (mTOR) and S6 kinase down-regulate phospholipase D2 basal expression and function. J. Biol. Chem. 285, 18991-9001.

Frondorf K, Henkels KM, Frohman MA, Gomez-Cambronero J. (2010) Phosphatidic acid is a leukocyte chemoattractant that acts through S6 kinase signaling. J. Biol. Chem. 285, 15837-47.

Henkels KM, Frondorf K, Gonzalez-Mejia ME, Doseff AL, Gomez-Cambronero J. (2011) IL-8-induced neutrophil chemotaxis is mediated by Janus kinase 3 (JAK3). FEBS Lett. 585, 159-66.

Peng H-J, Henkels KM, Mahankali M, Dinauer MC and Gomez-Cambronero J. (2011) Evidence for two CRIB domains in Phospholipase D2 (PLD2) that the enzyme uses to specifically bind to the small GTPase Rac2. J. Biol. Chem. 286, 16308-20.

Henkels KM, Farkaly T, Mahankali M, Segall JE and Gomez-Cambronero J. (2011) Cell Invasion of highly metastatic MTLn3 cancer cells is dependent on Phospholipase D2 (PLD2) and Janus Kinase 3 (JAK3). J. Mol. Biol. 408, 850-62.

Mahankali M, Peng H-J, Cox D and Gomez-Cambronero J. (2011) The mechanism of cell membrane ruffling relies on a Phospholipase D2 (PLD2), Grb2 and Rac2 association. Cell Signal. 23, 1291-8.

Peng J-H, Henkels KM, Mahankali M, Marchal C, Bubulya P, Dinauer MC and Gomez-Cambronero J. (2011) The dual effect of Rac2 on PLD2 regulation that explains both the onset and termination of chemotaxis. Mol. Cell. Biol. 31, 2227-40.

Gomez-Cambronero J. (2011) The exquisite regulation of PLD2 by a wealth of interacting proteins: S6K, Grb2, Sos, Wasp and Rac2 (and a surprise discovery: PLD2 is a GEF). Cell Signal. 23, 1885-95.

Kantonen S, Mahankali M, Hatton N, Henkels, K, Park H, Cox, D and Gomez-Cambronero J. (2011) A novel PLD2-Grb2-WASp protein heterotrimer regulates leukocyte phagocytosis in a two-step mechanism. Mol. Cell. Biol. 31, 4524-37.

Mahankali M, Peng H-J, Henkels KM, Dinauer MC and Gomez-Cambronero J. (2011) Phospholipase D is a GTP exchange factor (GEF) for the GTPase Rac. Proc Natl Acad Sci U S A. 108, 19617-22.

Di Fulvio, Frondorf K, Henkels, K, Grunwald Jr WC, Cool D and Gomez-Cambronero J. (2012) Phospholipase D2 (PLD2) shortens the time required for myeolid leukemic cell differentiation: Mechanism of action. J. Biol. Chem. 287, 393-407.

Gomez-Cambronero J, Allen L-A H, Cathcart MK, Justment L, Kovacs EJ, McLeish KR and Nauseef WM. (2012) How to Write Your First Grant Proposal: An Educational Workshop Organized by the Society for Leukocyte Biology. Nature Immunology. 13, 105-8.

Gomez-Cambronero J and Henkels K. (2012) Cloning of PLD2 from baculovirus for studies in inflammatory responses. Methods Mol. Biol. In Press.