APPLING LAB

HOME OF THE FOLIC ACID RESEARCH TEAM

 Publications

Protein Structure
and Enzymology

Knock-out Mouse Models
and Developmental Biology

Molecular Biology

 Projects
 Lab Stuff  Sequences
 People

In the News

 


Recent Publications:

Momb, J., and Appling, D.R. (2014) "Mitochondrial One-Carbon Metabolism and Neural Tube Defects" Birth Defects Research Part A: Clinical and Molecular Teratology 100, 576-583 [BDRA Online]

Shin, M., Bryant, J.D., Momb, J., and Appling, D.R. (2014) “Mitochondrial MTHFD2L is a Dual Redox Cofactor Specific Methylenetetrahydrofolate Dehydrogenase/Methenyltetrahydrofolate Cyclohydrolase Expressed in Both Adult and Embryonic Tissues” J. Biol. Chem. 289, 15507-15517 [JBC Online]

Momb, J. Lewandowski, J.P., Bryant, J.D., Fitch, R., Surman, D.R., Vokes, S.A., and Appling, D.R. (2013) “Deletion of Mthfd1l Causes Embryonic Lethality and Neural Tube and Craniofacial Defects in Mice” Proc. Natl. Acad. Sci. USA 110, 549-554 [PNAS Online]

Bolusani, S., Young, B.A., Cole, N.A., Tibbetts, A.S., Momb, J., Bryant, J.D., Solmonson, A., and Appling, D.R. (2011) "Mammalian MTHFD2L encodes a mitochondrial methylenetetrahydrofolate dehydrogenase isozyme expressed in adult tissues" J. Biol. Chem. 286, 5166-5174 [JBC Online]

Tibbetts, A.S. and Appling, D. R. (2010) “Compartmentalization of Mammalian Folate-Mediated One-Carbon Metabolism” Ann. Rev. Nutrition 30: 57-81. [ARN Online]

Textbook: Mathews, C.K., van Holde, K.E., Appling, D.R., and Anthony-Cahill, S. (2013) Biochemistry, 4th edition. Pearson Canada


Lab Stuff

 Frozen Yeast Stocks  
   
     

 

 


Current Lab Members:

applab2010

Left to right: Anne Tibbetts, Nicola Cole, Josh Bryant, Jane Shin, Paul Gonzalez, Mitzi Aguilar, Ming Chung, Jessica Momb

Dean Appling

Josh Bryant Graduate Student (B.S. Chemistry 2009 Hardin-Simmons Univ., Abilene TX)

Jessica Momb Postdoctoral Associate (Ph.D. in Biochemistry 2009 UT Austin

Min Hye Shin Graduate Student (B.S. Food Science 2006, Korea Univ.; M.S. Food Biotechnology 2008, Korea Univ.)

 

Former Students and Postdoctoral Fellows and Collaborators

Charles Barlowe (B.S. Chemistry 1983 William and Mary) Received Ph.D. in Chemistry (Biochemistry) 1990. Dissertation: "Investigation of the Enzymology and Metabolic Function of C1-Tetrahydrofolate Synthase". Charlie is a Full Prof. and Chair of the Department of Biochemistry at Dartmouth Medical School.

Andrew Breksa (B.S. Chemistry 1994 Brigham Young University) Received Ph.D. in Chemistry (Biochemistry) 1998. Dissertation: "A Multidisciplinary Approach to the Examination of the Catalytic Mechanism, Kinetic Mechanism, and Metabolic Role of the NAD-Dependent 5,10-Methylenetetrahydrofolate Dehydrogenase (MTD1p) in Saccharomyces cerevisiae". Andrew is a Research Chemist at the USDA Agricultural Research Service Western Regional Research Center in Albany, CA.

Swetha Bolusani (B. Tech. Chem. Eng. 2003 Jawaharlal Nehru Tech. Univ., Hyderbad, India; M.S. Chem. Eng. 2005 Louisiana Tech, Ruston LA) Received M.S. in Biochemistry December 2009. Thesis: "Identification and Characterization of the Rat Methylene-THF Dehydrogenase Encoded by the MTHFD2L Gene".

Sherwin Chan (M.S. Physiology and Cell Biology 1999 University of Alberta, Edmonton Alberta, Canada) Received Ph.D. in Biochemistry 2003. Dissertation: "Function and Regulation of Two Methylenetetrahydrofolate Reductase isozymes in Saccharomyces cerevisiae". Sherwin did a postdoctoral fellowship in the laboratory of Dr. Gerald Fink, The Whitehead Institute, MIT. He is now doing patent work for the law firm of Fish & Richardson in Boston, MA.

Leon Fernando Garcia Martinez (B.S. Chemistry 1986 Inst. Tecnologico y de Estudios Superiores de Monterrey, Mexico) Received Ph.D. in Biochemistry 1993. Dissertation: "Studies on the Compartmentation of Folate-Mediated One-Carbon Metabolism". Leon is Associate Director of Research at Alder Biopharmaceuticals in Bothell, WA.

Cristiana Garofalo (Ph.D. Chemistry University of Camerino, Italy) Postdoctoral Associate 2002-2004. Cristiana is now on the Faculty of Agronomy in the Dipartimento di Scienze Alimentari, Agro-Ingegneristiche, Fisiche, Economico-Agrarie e del Territorio (SAIFET) of the Università Politecnica Marche at Ancona, Italy.

Jennifer Herrera (Ph.D. Biochemistry 1999 Rice University) Postdoctoral Associate 2000-2001

Will Holmes (B.S. Zoology, 1990 University of Louisville) Received Ph.D. in Biochemistry 2001. Dissertation: "Characterization of Yeast Methenyltetrahydrofolate Synthetase and study of the Requirement for Formylation of Initiator tRNAfmet in Yeast Mitochondria". Will works in the laboratory of Dr. Hong Ye in the Brown Cancer Center at the Univ. of Louisville, Louisville, KY.

Katherine Howard (B.S. Marine Science 1985 Texas A&M Galveston) Received Ph.D. in Biochemistry 1994. Dissertation: "Rat C1-Tetrahydrofolate Synthase Gene: Characterization of the Promoter and Intron/Exon Boundaries". Kathy is an Assistant Professor of Biomedical Sciences at the University of Nevada Las Vegas UNLV School of Dental Medicine.

Stephanie Joslin (B.S. Microbiology; B.A. Plan II, 2003 UT Austin) Stephanie is a graduate student at the University of Texas-Southwestern Medical School in Dallas.

Evdokia Kassini Kastanos (B.A. Biochemistry 1990 UT-Austin) Received Ph.D. in Biochemistry 1996. Dissertation: "The Roles of Serine Hydroxymethyltransferase and Methylenetetrahydrofolate Reductase Isozymes in One-Carbon Metabolism". Evdokia is an Assistant Professor in the Department of Life and Health Sciences, University of Nicosia in Cyprus.

Tim Kirksey (B.S. Chemistry 1987 University of Oklahoma) Received Ph.D. in Biochemistry 1994. Dissertation: "Investigation of the Function of Asp449 in C1-Tetrahydrofolate Synthase". Tim is a Consultant Analyst Programmer in Discovery IT at GlaxoSmithKline R&D, North Carolina.

Gisela Kramer (Ph.D. Zoophysiology 1967 Univ. of Kiel, Germany). Gisela came to UT in 1974 to work with Boyd Hardesty, and was a Senior Research Scientist in the Department of Chemistry and Biochemistry. After Hardesty retired, Gisela and I collaborated for 8 years on mitochondrial protein synthesis. Gisela retired in 2008 and returned to her home in Lübeck, Germany.

David (Changkeun) Lee (M.S. Chemistry 2000 Seoul National University, Korea) Received Ph.D. in Biochemistry August 2008. Dissertation: "Biochemical and Genetic Studies of Mitochondrial Protein Synthesis in Saccharomyces cerevisiae: Characterization of the AEP3 and TRM5 Gene Products". David is now a Postdoctoral Fellow with Youxing Jiang in the Dept. of Physiology, UT Southwestern Medical Center at Dallas.

Stephanie Muga (B.S. Biology 1986 University of North Carolina) Received Ph.D. in Biochemistry 1995. Dissertation: "Gene Expression of the Rat C1-Tetrahydrofolate Synthase in Regenerating Liver and Developing Rat Liver and Rat Kidney". Stephanie is an Instructor of Biology in the Dept. of Biology and Geology at the Univ. of S. Carolina Aiken.

Laura Pasternack (B.S. Biochemistry 1988 University of New Mexico) Received Ph.D. in Biochemistry 1994. Dissertation: "13C NMR Analysis of Folate-Mediated One-Carbon Metabolism in Yeast". Laura is Assoc. Director of the NMR Facility, The Scripps Res. Institute, La Jolla, CA.

Schuyler Pike (B.A. Biology 1992 Cornerstone University, Grand Rapids MI) Received Ph.D. in Biochemistry December 2008. Dissertation: "Role of Mitochondrial C1-Tetrahydrofolate Synthase in embryogenesis". Sky is now a postdoctoral researcher in the laboratory of Dr. Norbert Kaminski in the Center for Integrated Toxicology at Michigan State University.

Priya Prasannan (B.S. Microbiology 1997 St. Aloysius College, India; M.S. Biochemistry 1999 Maharaja College, India) Received Ph.D. in Biochemistry December 2006. Dissertation: " Characterization of a Gene Encoding the Human Mitochondrial C1-Tetrahydrofolate Synthase and Submitochondrial Localization of the Protein".

Rhonda Raymond (B.S. Physical Science Education, 1994 University of New Mexico) Received Ph.D. in Biochemistry August 2000. Dissertation: "Isolation and Characterization of Two Genes Encoding Methylenetetrahydrofolate Reductase Isozymes from Saccharomyces cerevisiae". Rhonda is back at UT as a Lecturer teaching undergraduate Biochemistry.

Anice Thigpen (B.S. Chemistry 1979 Louisiana College) Received Ph.D. in Biochemistry 1990. Dissertation: "Rat C1-Tetrahydrofolate Synthase: Isolation of the Complementary DNA and Gene, Tissue-Specific Levels of the Messenger RNA, and Expression of the Protein in Yeast".

Anne Tibbetts (B.S. Chemistry/Biochemistry, 1993 Boise State University) Received Ph.D. in Biochemistry August 1999. Dissertation: "Isolation and characterization of two Saccharomyces cerevisiae AICAR transformylase/IMP cyclohydrolase isozymes". Anne is now the Director for Postgraduate Education in the College of Natural Sciences at UT..

Elizabeth Trimmer (Ph.D. Biochemistry Michigan) Elizabeth is an Assoc. Professor of Chemistry at Grinnell College, Iowa. Elizabeth spent the 2007-2008 academic year on a joint sabbatical with Ken Johnson and me.

Bert Vick (B.S. Biochemistry 2000 UT-Austin; Ph.D. Univ. of California - Berkeley) Bert is cofounder of Aurora Biofuels - an algal bio-energy start-up in the Bay Area.

Addie Walkup (B.S. Biochemistry 2002 McMurry University, Abilene TX) Received M.S.in Biochemistry August 2005. Thesis: "Characterization of the 10-Formyl-Tetrahydrofolate Synthetase Activity of the Human Mitochondrial C1-Tetrahydrofolate Synthase".

Russell Ward (B.S. Biochemistry, 1998, UT-Austin). Former undergraduate researcher and Research Assistant, Russell earned an M.D. at Texas A&M University Medical School. Russell is now doing Musculoskeletal Oncology surgery at Scott and White Memorial Hospital in Temple TX.

Mary West (B.S. Biochemistry 1985 University of Dallas) Received Ph.D. in Biochemistry 1996. Dissertation: "The Metabolic Role of the NAD-dependent 5,10-Methylenetetrahydrofolate Dehydrogenase in Saccharomyces cerevisiae". Mary is a Research Scientist with Odyssey Pharmaceuticals, San Ramon, CA.

Yakov Woldman (Ph.D. Biochemistry, 1993, Inst. of Bioorganic Chemistry, Novosibirsk, Russia). Postdoctoral Associate 1994-2000. Yakov is an Assoc. Professor in the Department of Chemistry at Valdosta St. University, Valdosta GA.

Liwen Zhang (B.S. Cell Biol and Genetics, 1988 Peking University) Received M.S. in Biochemistry from UT in August 1993 Thesis: "Intron-exon organization of rat C1-THF synthase". Liwen earned her Ph.D. from Rutgers and is in the Dept. of Cardiovascular Diseases, Merck Research Laboratories, Rahway, NJ.


 

Projects

The lab has several ongoing projects, all more or less concerned with the cellular organization of folate-mediated one-carbon metabolism. We are trying to understand how metabolic pathways are structured and compartmentalized within eukaryotic cells. Some of these studies use the yeast Saccharomyces cerevisiae as a model system while others rely on whole animals (rats and mice) and mammalian cell lines. We typically focus on specific enzymes within a particular pathway, studying their biochemistry, structure and interactions with other proteins. Techniques include classical protein chemistry, recombinant DNA and molecular genetics, metabolic engineering, knockout mouse models, and NMR to follow the metabolism.Currently we are focusing on:


Lab Protocols
High Efficiency Yeast Transformation Tetrahydrofolate Prep

Methylene-THF
dehydrogenase assay

(6-R,S)Methyl-THF Prep
Cobalamin-Dependent Methionine Synthase Assay Methenyl-THF cyclohydrolase assay 10-formyl-THF
synthetase assay
 

Last update: 23 September, 2014