Curriculum vitae david andrew johnson

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Department of Biomedical Science

Professor of Biochemistry and Molecular Biology

J. H. Quillen College of Medicine

East Tennessee State University

Johnson City, TN 37614-0581david head bow tie 2008

Phone (423) 439-2027

FAX (423) 439-2030



1967 B.S. in Chemistry, ACS approved, University of Memphis

1973 Ph.D. in Chemistry, University of Memphis

1990-Present Professor of Biochemistry, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, Tennessee

1993-95 Interim Chair of Biochemistry, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, Tennessee (7/1/93-9/30/95)

1995 Adjunct Professor of Biology, College of Arts and Sciences, ETSU

1984-1990 Associate Professor of Biochemistry, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, Tennessee

1978-1984 Assistant Professor of Biochemistry, Quillen-Dishner College of Medicine, East Tennessee State University, Johnson City, Tennessee

1976-1978 Assistant Biochemist, Department of Biochemistry, University of Georgia, Athens, Georgia

1974-1976 National Heart and Lung Institute Postdoctoral Fellow, Department of Biochemistry, University of Georgia, Dr. James Travis - Mentor

1973-1974 Research Associate, Biochemistry, University of Georgia, Dr. James Travis

1972 & 1973 Instructor of Biochemistry, Chemistry Department, University of Memphis,

(Summers) Memphis, Tennessee

1967-1973 Graduate Teaching Assistant, Chemistry Department, University of Memphis, Memphis, Tennessee

1967 & 1968 Chemist, Antigen Preparation Laboratory, TB Branch,

(Summers) National Center for Disease Control, Atlanta, Georgia
1989 Visiting Professor, University of British Columbia, Vancouver,Canada.

1987 East Tennessee State University Foundation Research Award for study of "The Biochemical Aspects of Pulmonary Diseases in Man". This is the highest recognition given for research each year. Selection is made by a committee of peers.
1974 NIH Post-doctoral Fellowship to study the structure and function of human alpha-1 proteinase inhibitor at the University of Georgia with Dr. J. Travis -July 1, 1974 - June 30, 1976.

2008 J. H. Quillen V. A. Medical center biomedical research corporation board of directors

2010 J.H. Quillen Dean’s Teaching Award for Basic Science
2000 Visiting Professor in Immunopharmacology at the University of Southampton for three months, August - October, in the laboratory of Dr. Andrew Walls.
1997 Methods in Molecular Biology Workshop - ETSU COM July 14-25, 1997

1991 Wellcome visiting professor in the Laboratory of Molecular Biophysics with Dr. Geoffrey Barton, University of Oxford, England, May and June, "Molecular Modeling of Mast Cell Tryptases", supported by a Burroughs Wellcome Fund Research Travel Grant - May 1 -June 30, 1991.

1990 Course on computer based multiple sequence alignment of DNA and proteins, Pittsburgh Super Computer Center - Aug 5-8; all expenses paid by the PSCC.

1985 Wellcome visiting scientist at Strangeways Research Laboratory with Dr. Alan Barrett, Cambridge, England, April 15 through July 15; "Human Mast Cell Tryptase", supported by a Burroughs Wellcome Fund Research Travel Grant


Research deals with proteolytic enzymes and their inhibitors, including protein purification, characterization, sequencing, structure function relationships, kinetics, and regulation. Particular emphasis is placed on the roles of proteases and inhibitors in the pathogenesis of human diseases. Work on the structure of human alpha-1-proteinase inhibitor led to the discovery of its active site and we showed that oxidation of a methionine residue in the inhibitory site of α1-PI caused the loss of elastase inhibitory activity. This finding resulted in the hypothesis that oxidants can lead to a lung-localized inhibitor deficiency. A deficiency of α1-PI, which normally inhibits neutrophil elastase, is known to cause emphysema. This led to studies of the reactions of ozone and nitrogen dioxide on the function of human α1-PI and the secretory leukocyte proteinase inhibitor found in human bronchial mucus, which also inhibits neutrophil elastase. Another long-standing project focuses on the structure and function of human mast cell tryptase, an unusual serine protease that is the most abundant protein in mast cells. Recently, tryptase has been shown to activate protease zymogens, suggesting a critical role in tumor growth and the pathogenesis of arthritis. Pichia pastoris has been used to expression of recombinant human mast cell and neutrophil serine proteases, as well as human enterokinase and C-reactive protein.

Current Committee Service:
Medical Student Admissions Committee

Medical Student Education Committee

M1/M2 Medical Student Curriculum Review - Chair


1. J. Travis, D. A. Johnson, and R. Pannell. Properties of Human Alpha-1-Antitrypsin. Bayer Symposium V, "Proteinase Inhibitors" (1974)
2. D. Johnson, R. Pannell, and J. Travis. The Molecular Stoichiometry of Trypsin Inhibition by Human Alpha-1-Proteinase Inhibitor. Biochem. Biophys. Res. Comm. 57. 584-589 (1974)
3. R. Pannell, D. Johnson, and J. Travis. Purification and Properties of Human Alpha-1-Proteinase Inhibitor. Biochemistry 13, 5439 (1974)
4. D. Johnson and J. Travis. Mechanism and Structure of Alpha-1-Proteinase Inhibitor. XXIIIrd Colloquium, "Protides of the Biological Fluids", ed. H. Peters, Vol. 23, 35-38, Pergamon Oxford (1975)
5. D. Johnson and J. Travis. Rapid Purification of Human Trypsin and Chymotrypsin I. Analytical Biochem. 72, 573-576 (1976)
6. R. Baugh, D. Garner, D. Johnson, and J. Travis. Regulation of the Leukocyte Proteinases by the Human Plasma Proteinase Inhibitors: Proteolysis and Physiological Regulation. Miami Winter Symposium, Vol. 11, ed. D. W. Ribbons and K. Brew, Academic Press, N.Y. 393 (1976)
7. J. Travis, J. Bowen, D. Tewksbury, D. Johnson, and R. Pannell. Whole Human Plasma: Isolation of Albumin and Fractionation of Albumin Depleted Plasma. Biochem. Journal 157, 301-306 (1976)
8. D. A. Johnson. Purification and Properties of Rabbit Trypsin. Biochim. Biophys. Acta, 452, 482-487 (1976)
9. D. A. Johnson and J. Travis. Human Alpha-1-Proteinase Inhibitor Mechanism of Action: Evidence for Activation by Limited Proteolysis. Biochem. Biophys. Res. Comm. 72, 33-39 (1976)
10. D. A. Johnson and J. Travis. Inactivation of Human Alpha-1-Proteinase Inhibitor by Thiol Proteinases. Biochem. Journal 163, 639-641 (1977)
11. J. Travis, N. Matheson, and D. A. Johnson. The Interaction of Alpha-1-Proteinase Inhibitor with Serine Proteinases. Chemistry and Biology of Thrombin, eds. R. Lundblad, J. Fenton, and K. Mann, p. 431-440, Ann Arbor Science Publishers, Ann Arbor, Mich. (1977)
12. A. B. Dudkiewicz, W. F. Lehnhardt, D. A. Johnson, J. Travis, and W. L. Williams. Rabbit Acrosin: Immunological Dissimilarity to Rabbit Trypsin. J. Exp. Zool. 203, 277-286 (1978)
13. J. Travis and D. Johnson. The Carboxy Terminal Sequence of Human Alpha-1-Proteinase Inhibitor. Biochem. Biophys. Res. Commun. 84, 219-224 (1978)
14. D. Johnson and J. Travis. Structural Evidence for Methionine at the reactive Site of Human Alpha-1-Proteinase Inhibitor. J. Biol. Chem. 253. 7142-7144 (1978)
15. J. Travis, R. Baugh, P. Giles, D. Johnson, J. Bowen, and C. Reilly. Human Leukocyte Elastase and Cathepsin G: Isolation, Characterization and Interaction with Plasma Proteinase Inhibitors. Neutral Proteases and Human Polymorphonuclear Leukocytes, eds. K. Havemann and A. Janoff. Urban and Schwarzenberg, Munich (1978)

16. D. Johnson and J. Travis. The Oxidative Inactivation of Human Alpha-1-Proteinase Inhibitor: Further Evidence for Methionine at the Reactive Center. J. Biol. Chem. 254, 4022-4026 (1979)
17. J. Travis, N. Matheson, D. Johnson, and K. Beatty. Human Alpha-1-Proteinase Inhibitor and Human Alpha-1-Antichymotrypsin: Properties and Mechanism Studies. The Chemistry and Physiology of the Human Plasma Proteins, ed. D. H. Bing, Pergamon Press Inc., N.Y., p. 343-352 (1979)
18. D. A. Johnson. Ozone Inactivation of Human Alpha-1-Proteinase Inhibitor. Am. Rev. Respir. Dis. 121, 1031-1038 (1980)
19. J. Travis and D. Johnson. Human Alpha-1-Proteinase Inhibitor. Methods in Enzymology, Vol. 80, ed. L. Lorand. Academic Press, N.Y. p. 754-765 (1981)
20. D. A. Johnson, B. Carter-Hamm, and W. Dralle. Inactivation of Human Bronchial Mucosal Proteinase Inhibitor by Pseudomonas aeruginosa Elastase. Amer. Rev. Respir. Dis. 126, 1070-1083 (1982)
21. D. A. Johnson. A Procedure for the Separation and Quantitation of Tryptophan and Amino Sugars on the Amino Acid Analyzer. Analytical Biochemistry, 130 475-480 (1983)
22. T. Tanaka, B. J. McRae, K. Cho, R. Cook, J. E. Fraki, D. A. Johnson, and J. C. Powers. Mammalian Trypsin-Like Enzymes. Comparative Reactivities of Human Skin Tryptase, Human Lung Tryptase and Bovine Trypsin with Peptide 4-Nitroanilide and Thioester Substrates. J. Biol. Chem. 258, 13552-13557 (1983)
23. M. C. McDaniel, M. A. Tucker, and D. A. Johnson. Production of Macrophage Activation Factors by Tryptic Cleavage of Calf Serum Proteins. Inflammation, 1, 339-345 (1983)
24. T. Smith, M. Hougland, and D. A. Johnson. Human Lung Tryptase: Purification and Characterization. J. Biol. Chem. 259, 11046-11051 (1984)
25. C. E. Smith and D. A. Johnson. Human Bronchial Leucocyte Proteinase Inhibitor: Rapid Isolation and Kinetic Analysis with Human Leucocyte Proteinases. Biochem. J. 225, 463-472 (1985)
26. D. A. Johnson and T. E. Cawston. Human Lung Mast Cell Tryptase Fails to Activate Procollagenase or Degrade Proteoglycan. Biochem. Biophys. Res. Comm. 132, 453-459 (1985)
27. J. Travis and D. Johnson. Inhibitors of Proteolytic Enzymes that Function in the Lung. In Mechanisms of Lung Injury. A Symposium Held at The Graduate Hospital in Philadelphia, Pennsylvania, Oct. 1 & 2, 1984. (T. P. Stein and G. Weinbaum, editors). George F. Stickely, Co. pp. 1-11 (1986)
28. R. W. Mason, D. A. Johnson, A. J. Barrett, and H. A. Chapman. Elastinolytic Activity of Human Cathepsin L. Biochem. 233, 925-927 (1986)
29. D. A. Johnson, R. S. Winters, T. Wooley, D. Graham, and F. W. Henderson. Ozone Effects on Alpha-1-Proteinase Inhibitor In Vivo: Blood Plasma Inhibitory Activity is unchanged. Exp. Lung Res. 11, 95-103 (1986)

30. D. A. Johnson, A. J. Barrett, and R. W. Mason. Cathepsin L. Inactivates α1-Proteinase Inhibitor by Cleavage in the Reactive Site Region. J. Biol. Chem. 261, 14748-14751 (1986)

31. C. E. Smith, M. S. Stack, and D. A. Johnson. Ozone Effects on Inhibitors of Human Neutrophil Proteinases. Archives Biochem. Biophys. 253, 146-155 (1987)

32. J. A. Cromlish, N. G. Seidah, M. Marcinkiewicz, J. Hamelin, D. A. Johnson, and M. Chretien. Human Pituitary Tryptase: Molecular Forms, NH2-terminal Sequence, Immunochemical Localization, and Specificity with Prohormone and Fluorogenic Substrates. J. Biol. Chem. 262, 1363-1373 (1987)
33. D. A. Johnson, G. Salvesen, M. A. Brown, and A. J. Barrett. Rapid Isolation of Human Kininogens. Thrombosis Res. 48, 187-193 (1987)
34. D. A. Johnson. Effects of Ozone and Nitrogen Dioxide on Human Lung Proteinase Inhibitors, Health Effects Institute Research Report No. 11, August 1987.
35. M. S. Stack, C. E. Smith, W. L. Dean, and D. A. Johnson. Bronchial Leukocyte Proteinase Inhibitor: Hydrodynamic Properties and Interaction with Alpha2-Macroglobulin-Bound Elastase. Archives Biochem. Biophys. 260, 400-407 (1988)
36. J. J. Ochino, R. T. Abboud, S. Lam, S. S. Johal, C. E. Smith, and D. A. Johnson. Bronchial Leukocyte Proteinase Inhibitor Levels in Bronchial Washings in Asthma Patients. Chest. 93, 1008-1013 (1988)
37. W. Müller-Esterl, D. Johnson, G. Salvesen, and A. J. Barrett. Human Kininogens, Purification, Fragmentation, and Isolation of the Major Fragments. In: Methods in Enzymology pp. 240-256, Vol. 163, DiSabato, G., ed. Academic Press, New York (1988)
38. C. E. Smith, P. R. Musich, and D. A. Johnson. Sodium Dodecyl Sulfate Enhancement of Quantitative Immunoenzyme Dot-Blot Assays on Nitrocellulose. Analytical Biochemistry 177, 212-219 (1989)
39. D. A. Johnson, M. W. Frampton, R. S. Winters, P. E. Morrow, and M. J. Utell. Inhalation of Nitrogen Dioxide Fails to Reduce the Activity of Human Lung Alpha-1-Proteinase Inhibitor. Am. Rev. Respir. Dis. 142:758-762 (1990)
40. D. A. Johnson, R. S. Winters, K. R. Lee, and C. E. Smith. Oxidant Effects on Rat and Human Lung Proteinase Inhibitors. Research Report (#37) to be published by the Health Effects Institute Project No. 85-4 (reviewed internally and externally by HEI). Library of Congress Cat. #WA 754 R432 (1990)
41. D.A. Johnson and G. J. Barton. Mast Cell Tryptases: Examination of Unusual Characteristics by Multiple Sequence Alignment and Molecular Modeling. Protein Science 1, 370-377 (1992)
42. W. Chu, D. A. Johnson, and P. R. Musich. Molecular Cloning and Characterization of Mouse Mast Cell Chymases. Biochim. Biophys. Acta 1121, 83-87 (1992)
43. C. E. Smith, H. S. Koren, D. E. Graham, and D. A. Johnson. Mast Cell Tryptase is Increased in the Nasal and Bronchoalveolar Lavage Fluids of Humans after Ozone Exposure. Inhalation Toxicology 5, 117-127 (1993)
44. C. P. Ebenbichler, Westervelt, C. Carillo, T. Henkel, D. Johnson, and L. Ratner. Structure-function Relationships of the HIV-1 Envelope V3 Loop Tropism Determinant: Evidence for Two Distinct Conformations. AIDS 7, 639-646 (1993)

45. D. B. Hood, Gettins, P. and D. A. Johnson. Reaction of Nitrogen Dioxide with Proteins; Effects on Activity and Immunoreactivity with Alpha-Proteinase Inhibitor and Implications for NO2 Mediated Polypeptide Degradation. Arch. Bioch. Biophys. 304, 17-26 (1993)

46. M. S. Stack and D.A. Johnson. Human Mast Cell Tryptase Activates Single Chain Urinary-Type Plasminogen Activator (Prourokinase). J. Biol. Chem. 269, 9416-9419 (1994)
47. R. S. Winters, B. Burnette-Vick, and D. A. Johnson. Ozone, but Not Nitrogen Dioxide, Fragments Elastin and Increases Its Susceptibility to Proteolysis Amer. J. Respir. Dis. Crit. Care Med. 150, 1026-31 (1994).
48. S. S. Little. and D. A. Johnson. Human Mast Cell Tryptase Isozymes: Separation and Examination of Substrate Specificity Differences. Biochem. J. 307, 341-346 (1995)
49. A. K. Addington and D. A. Johnson, Inactivation of Human Lung Tryptase: Evidence for a Re-activatable Tetrameric Intermediate and Active Monomers. Biochemistry 35, 13511-13518 (1996).
50. D.A. Johnson Tryptase. In Handbook of Proteolytic Enzymes. (In book form and as a searchable CD ROM) (A.J. Barrett, F. Woessner, and N. Rawlings, eds.) Academic Press (1998).

51. V.A. Thomas, C. J. Wheeless, M. S. Stack and David A. Johnson Human Mast Cell Tryptase Fibrinogenolysis: Kinetics, Anticoagulation Mechanism and Cell Adhesion Disruption. Biochemistry 37, 2291-2298 (1998).

52. Andrew L. Niles, Mark Maffit, Mary Haak-Frendscho, Christine J. Wheeless, and David A. Johnson, Recombinant Human Mast Cell Tryptase Beta: Stable Expression in Pichia pastoris and Purification of Fully Active Enzyme. Biotechnol. Appl. Biochem. 28:125-131 (1998).

D.S. Jackson, S.A. Fraser, L-M. Ni, C-M. Kam, U. Winkler, D.A. Johnson, C.J. Froelich, D Hudig, and J.C. Powers. Synthesis and Evaluation of Diphenyl Phosphonate Esters as Inhibitors of the Trypsin-like Granzymes A and K and Mast Cell Tryptase. J. Med. Chem.18:22892301 (1998).

Fusun Kilic, David A. Johnson and Michael Sinensky Subcellular localization and partial purification of prelamin A endoprotease: an enzyme which catalyzes the conversion of farnesylated prelamin A to mature lamin A. FEBS Letters 450:61-65 (1999).
55. Rosealee Smith, Alison Jenkins, Afrodite Lourbakos, Philip Thompson, Vanitha Ramakrishnan, Jim Tomlinson, Usha Deshpande, David A. Johnson, Roy Jones, Eleanor J. Mackie and Robert N. Pike Evidence for the activation of PAR-2 by the sperm protease, acrosin: expression of the receptor on oocytes. FEBS Letters 484; 285-290 (2000).
56. Krishnaswamy G, Kelley J, Johnson D, Youngberg G, Stone W, Huang SK, Bieber J, Chi DS. The human mast cell: functions in physiology and disease. Front Biosci 2001 Sep 1; 6:D1109-D1127.
57. Krishnaswamy G, Hall, K., Youngberg, G., Hossler, F., Johnson D, Block, W.A., Huang, S-K., Kelley, J., and Chi D.S. Regulation of eosinophil-active cytokine production from human cord blood-derived mast cells. Journal Interferon and Cytokine Research 22:379-388 (2002).
58. Lockhart BE, Vencill JR, Felix CM and Johnson DA. Recombinant Human Mast Cell Chymase: Improved Expression in Pichia pastoris and Purification of Highly Active Enzyme Biotechnology and Applied Biochemistry 41, 89–95 (2005).

59. Johnson DA Human Mast Cell Proteases: Activity Assays Using Thiobenzyl Ester Substrates chapter in “Protocols in Molecular Biology: The Human Mast Cells.” G. Krishnaswamy and D. Chi eds. Clifton, NJ 315, 193-202, 2006.

60. Burrows, DL, Nicolaides, A, Rice, PJ, Dufforc, M, Johnson, DA and Ferslew, KE. Papain: A Novel Urine Adulterant. Journal of Analytical Toxicology 29:275-295 (2005).
61. Coffman, Lan; Brown, Julie; Johnson, David; Parthasarathy, Narayanan; D'Agostino Jr., Ralph; Lively, Mark; Hua, Xiaoyang; Tilley, Stephen; Muller-Esterl, Werner; Willingham, Mark; Torti, Frank; Torti, Suzy. Cleavage of High Molecular Weight Kininogen by Elastase and Tryptase is Inhibited by Ferritin. American Journal of Physiology 294, L505-515, 2008.
62. Smita Negi, Ahmad Halawa, David S. Chi, Christopher Miller, Fred Hossler, George Youngberg, David A. Johnson and Guha Krishnaswamy MAST CELL REGULATION OF CARDIOVASCULAR INFLAMMATION I: COGNATE AND NON-COGNATE INTERACTIONS In: Mast Cells and Cardiovascular Disease.  Ed. Joseph J, Nova Science Publishers, Hauppauge, NY 2009 accepted 3/10/2010
63. Sanjay K. Singh, Avinash Thirumalai, David J. Hammond, Jr., Michael K. Pangburn, Vinod K. Mishra, David A. Johnson, Antonio E. Rusiñol, and Alok Agrawal Exposing a hidden functional site of C-reactive protein by site-directed mutagenesis J.Biol.Chem. 287, 3550–3558, 2012.
64. Eliot T. Smith and David A. Johnson, Human Enteropeptidase Light Chain: Bioengineering of Recombinants and Kinetic Investigations of Structure and Function Protein Science 22, 577-585, 2013.
65. Expression of recombinant human mast cell chymase with Asn-linked glycans in glycoengineered Pichia pastoris. Smith ET, Perry ET, Sears MB, Johnson DA. Protein Expr Purif. 2014 Oct;102:69-75. doi: 10.1016/j.pep.2014.08.005. Epub 2014 Aug 12.

1967-1973 University of Memphis, undergraduate and graduate laboratories; as an instructor I taught the entire biochemistry course (lectures and labs) for senior undergraduates and biology graduate students for two summer terms (1972 & 1973).

1973-1978 University of Georgia, direction and supervision of graduate students, occasional lectures in biochemistry classes
1978-present Medical Biochemistry, East Tennessee State University, Protein structure and function and amino acid metabolism.
1979-1995 Protein Structure and Function a graduate course (formerly called Enzymology) taught in odd numbered years with Dr. M.L. Ernst-Fonberg
1998 Structure and Dynamics of Biological Macromolecules a new graduate course taught with Drs. Robinson, Ernst-Fonberg and Sinensky, responsible for ~1/3 of the course.
1985-1993 Course Director, Medical and Graduate Biochemistry, ETSU, responsible for curriculum design and implementation
2001 Course Director, Biomedical Sciences 4 Graduate Course; BIOM 5040/6040 Fundamentals of Biomedical Research IV Cell and Organ Interactions Credit: 3 semester hours Spring semester, last 7 weeks (6 ‘hours' per week)
2003-5 Course director of Proteins and Proteomics a graduate course on proteins and their structure and function
2007 Course Director for inaugural Pharmacy Biochemistry; taught 25% of first 2 courses
Ph.D. Dissertation Director: Students and their current positions.

Timothy J. Smith (Ph.D. 1985); Chief Executive Officer, BioSignia, 100 Europa Dr., Chapel Hill, NC 27514 (919) 933-2021. BioSignia markets a proprietary biomarker based technology for the prediction of chronic degenerative diseases and related crisis events.

Craig E. Smith (M.S. 1986 & Ph.D. 1991) VP for Research ThermoFisher.

Darryl B. Hood (Ph.D. 1991); Assoc. Professor, Vice Chair Pharmacology, 1005 D.B. Todd Blvd., Meharry Medical College, Nashville, TN 37208, (615) 327-6358. PI of an NIH grant to study the reaction of NO2 with proteins, a continuation of his PhD dissertation research. He was our first minority Ph.D. graduate.

Susan S. Little (Ph.D. 1993); Following graduation she took a Post-doctoral position with Dr. Louis Hersh, Chair of Biochemistry, Univ. of Kentucky, Lexington, KY After two years with Hersh she moved to Physiology with Dr. Getchell for two years, worked in Atlanta 2 years, Nuclear Fuels in Erwin, TN 1 year, moved back to Knoxville in 2007.

Brent Lockhart (Ph.D. May 2008) Expression, Purification and Characterization of the Mast Cell Proteases Chymase and Cathepsin G; a faculty member at Northeast State and co-founder of Proteogenesis biotech company in JC.

M.S. Thesis Director: Students and their current positions

M. Sharon Stack, Ph.D. (M.S. 1986); Prof. and Vice Chair of Pathology, University of Missouri Medical School, Columbia, MO.

David Brown (M.S. 1987); Armed Services Medical School, Bethesda, (MD 1994), Army Medical.

Alison Babelay (2006-07) MS student from Furman home in Oak Ridge, went to U. South Carolina medical school in Columbia, SC 2007; worked on CatG expression in Pichia.

Eliot Smith (2007-present) MS student from UT Knoxville working on the expression of codon optimized neutrophil Cathepsin G and elastase in Pichia pastoris; also expressed human enterokinase and CRP.
Medical and Undergraduate Students Receiving Research Experience in My Laboratory:

Ricky A. Thompson, M.D. (1986-87 summers) Medical Student went to a Residency in Pathology at Duke Univ., Current position unknown

Kathryn Kalbfleisch (1992 and 1993, summers), (1997 M.D. Graduate) (Resident position unknown)

Tara Miller (1995-96) Medical Student M.D. 2000, Resident position unknown

Vanessa Thomas (Fitsanakis) (1997-1998) Milligan College undergraduate, Ph.D. in molecular pathology at Vanderbilt.

Sarina Brown (Summer 1998) McNair Scholar Program

Andrenette Fleming (Spring 1998) Six week Senior Rotation in Biochemistry Research working on the production of Tryptase site-directed mutants in Pichia, went on to an Ob/Gyn Residency at Robert Wood Johnson Medical School, Camden, NJ.

Christopher Trabue (1999) Summer Research on tryptase in atherosclerotic plaques supported by the ETSU Cardiovascular Research Institute.

Andrew Hull (Spring 2001) Milligan College Undergraduate; now a practicing Physician’s Assistant.

Susannah Johnson (Summer 2001) Converse College Undergraduate joint project with Dr. G. Krishnaswamy, IM. (2010 M. D. Graduate).

Jessica Vencill Hankins (2003-4) Undergraduate Biology major at ETSU; first place in 2004 Southern Appalachian Research Forum

Melody McVey (2006-2008) ETSU Chemistry Major and Honors College worked on Cathepsin G codon optimization and Western blotting.

Jessica Pugh (2007-present) Pre-Med Med Honors College working on expression of neutrophil elastase in E. coli.

Haley Klimecki (2008-2009) Pre-Med Med Honors Chemistry working on expression of human neutrophil elastase in K. lactis: Received a Student Faculty Collaborative Grant in 2009.

Michael Bradfield (2008-2009) Former Peace Corp taking pre-med fast track course at ETSU; Pepperdine grad., work study student helped with making reagents and media.

Karie Hodges (2009-2012) First Place in the Biomedical Sciences Undergraduate Division at the 2011 Southern Appalachian Research Forum and second place in 2012

Mallory McGhee (2010-2011)

Branson Mauck (2011-2012)

Evan Perry – undergraduate (2011-Present)

Dustin Wood – undergraduate (2011-Present)

Meagan Sears – undergraduate (2012-present)

1975-present American Association for the Advancement of Science

1978-1997 American Thoracic Society

1980-2010 Sigma Xi

1982-present American Society for Biochemistry and Molecular Biology

1991-present The Protein Society

1994-present American Chemical Society


1976-1978 Proteinase Inhibitors of Human Lung and Leukocytes. National Heart, Lung and Blood Institute Young Investigator Award, Univ. of Georgia - $29,555/2 years.
1979-1981 Endogenous Proteolytic Enzymes and Inhibitors of the Lung. The Council for Tobacco Research U.S.A. Inc. - $101,000
1983-1986 Effects of O3, NO2, and SO2 on Human Lung Proteinase Inhibitors. The Health Effects Institute - $173,206
1985 Human Lung Mast Cell Tryptase. Travel Grant for 3 months work at Strangeways Research Laboratory, Cambridge, England, Burroughs Wellcome Fund - $4,800
1985-1988 Oxidant Effects on Rat Lung Proteinase Inhibitors. The Health Effects Institute - $224,709
1987-1988 Roche Biomedical Laboratories, Allergy Test Development - $3,000
1987 Sponsor of Mr. Ricky Thompson's Southern Medical Association Grant - $1,350
1987-1990 Sponsor for Mr. Darryl B. Hood's NIH MARC Predoctoral Fellowship - $28,269
1989-1992 Human Lung Mast Cell Tryptase - NIH Area Grant R15 HL42623 - $97,819
1990 Sick Building Syndrome - Environment Protection Agency - $4,000
1990-1993 Effects of O3 and NO2 on Human Lung Proteins - NIH R01 ES04999 $388,704/3 years

1991 Molecular Modeling of Mouse Mast Cell Tryptases travel grant for 2 months work at the Laboratory of Molecular Biophysics, University of Oxford (May and June), Burroughs Wellcome Fund, $6,500

1992 Grant of "Hyperchem" software for molecular modeling from Autodesk Inc.- $3,000
1993 Burroughs Wellcome International Visiting Professorship - Brought Prof. John Kay, Cardiff, Wales to ETSU - $3,053
1993 Sponsor of Ms. Kathryn Kalbfleisch’s Southern Medical Association Grant - $2,500
1994-96 “The tetrameric structure of human lung tryptase and the role of heparin.” (To study the kinetics of tryptase inactivation and structural changes.) ARRIS Pharmaceuticals post-doctoral salary and supplies. This positioned was filled by Dr. Adele Mountz Addington a graduate of Virginia Tech - $46,667/16 months.
1995-97 “Human Lung Mast Cell Tryptase: Heparin Binding and Fibrinogen Cleavage” American Heart Association, Tennessee Affiliate $25,000/year
1996-97 “Thrombolysis by Human Mast Cell Tryptase” ETSU Cardiovascular Research Institute $5,000.
1997-1998 “Expression and Purification of Human Mast Cell Tryptase.” ETSU Research Development Committee Grant $5,200.
1997-1999 “Expression and Purification of Recombinant Human Mast Cell Tryptase” Promega Corp. Support of supplies - $4,000

  1. “A Cyclodiene-induced Binding Protein” NIEHS R15ES08817; Dr. Larry Curtis, Environmental Health was the original PI; ($96,833 for 1997-2000) and I took over when he moved to Oregon State University in July of 2000 until the grant ended in December of 2000.

    1. “Recombinant Human Mast Cell Tryptases”, NIAID, NIH R15 AI45549, $126,954.

2007-2011 “Human Cathepsin G: Expression, C-Terminal Processing and Dual Specificity” National Heart Lung and Blood Institute 1R15HL091770 $210,230.

2011 Patent disclosures on the expression of recombinant human C-Reactive Protein in Pichia pastoris

2011 Patent disclosures on the expression of recombinant human neutrophil elastase in Pichia pastoris

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