Becker Lab

Donald Becker, Ph.D.
Professor
Redox Biology Center

Department of Biochemistry

University of Nebraska
N258 Beadle Center
Lincoln, NE 68588-0664
Office (402)472-9307
Email  dbecker3@unl.edu

EDUCATION

B.A. Bethel College - Chemistry (1989)
Ph.D. University of Minnesota, Twin Cities - Biological Chemistry (1994)
Post-Doctoral University of Minnesota, Twin Cities - Biological Chemistry (1994-1995)
Post-Doctoral University of Nebraska-Lincoln - Enzyme Mechanisms (1995-1998)

 

Research Interests:

proline metabolism, redox enzymology, redox regulation of proteins and gene expression, substrate channeling in enzymes

Research Overview

The amino acid proline has multifaceted roles including carbon and nitrogen flux, protein synthesis, osmolyte balance, and stress protection. There is growing evidence that proline metabolism can influence cell survival outcomes in microorganisms, plants, and animals. Proline effects diverse signaling pathways by the generation of reactive oxygen species (ROS) due to proline oxidation being coupled to the respiratory electron transport chain and contributing to cellular bioenergetics. Overall proline has become a very important metabolite that is thought to be involved in many cellular processes that impact human health and disease.

The overall goal of our research is to understand the mechanisms of proline metabolic enzymes and how proline metabolism impacts stress response and the intracellular redox environment. All organisms convert proline to glutamate in two enzymatic steps. In the first step, proline is oxidized to ∆1-pyrroline-5-carboxylate (P5C) by the flavin-dependent enzyme PRODH. P5C is then hydrolyzed nonenzymatically to glutamic semialdehyde (GSA), which is oxidized to glutamate by the NAD dependent enzyme, P5C dehydrogenase (P5CDH). In Gram-negative bacteria, the PRODH and P5CDH domains are fused onto the same polypeptide called the proline utilization A (PutA) protein. Proline biosynthesis from glutamate involves three enzymatic steps.  The initial two steps are catalyzed by g-glutamyl kinase (GK) and g-glutamyl phosphate reductase (GPR). GK generates g-glutamyl phosphate, which is then reduced by GPR to produce GSA.  In bacteria and lower eukaryotes such as yeast, GK and GPR are discrete monofunctional enzymes.  In animals and plants, the GK and GPR domains are fused together into the bifunctional enzyme P5C synthase (P5CS). After GSA cyclizes to P5C, P5C is reduced to proline by P5C reductase (P5CR).

 

Lab Members

Xinwen Liang

Research Technologist/Senior Scientist


Research Project
:

Proline biosynthesis and protective mechanisms of stress

E-mail: Xinwen

 Julia Zhang                                                                                                 Gong Yushun

Postdoctoral Associate                                                                       Visiting Scholar; Associate Professor, Hunan                                                                                                                                                                             Agricultural University

                                    
Research Project:                                                                                   Research Project:

Proline metabolic redox signaling;                                                  Proline biosynthesis and stress in C. elegans
enzymology of proline metabolism                                                                                                        
                                                                                                                     E-mail: Gong
E-mail: Julia                                                                                                                      

Shelbi Christgen                                                                                      Sagar Patel

Graduate Student                                                                                 Graduate Student

                                        
Research Project:                                                                                  Research Project: 

Multifunctional mechanisms of PutA; substrate                       Enzymology studies of proline metabolism
channeling; proline in microbial pathogenesis
                                                                                                                    E-mail: Sagar
E-mail: Shelbi                                                                                        

Joshua Floth                                                                                               Jacob Wilkinson  

Undergraduate Researcher                                                             Undergraduate Researcher


Research Project:                                                                                  Reserach Project:

Catalytic mechanisms of proline                                                    Crystallography of reduced PutA enzymes
dehydrogenase; substrate channeling
between enzyme pairs                                                                        E-mail: Jacob

E-mail: Josh                                                                                                                                                         

 

PubMed Articles  

Recent Publication List:

Zhang, L. and Becker, D.F. (2015) Connecting Proline Metabolism and Signaling Pathways in Plant Senescence Front Plant Sci 6:552

Sanyal, N.; Arentson, B.W.; Luo, M.; Tanner, J.J., Becker, D.F. (2015) First Evidence for Substrate Channeling Between Proline Catabolic Enzymes: A Validation of Domain Fusion Analysis for Predicting Protein-Protein Interactions J Biol Chem 290(4): 2225-34

Zhang, L.; Alfano, J.R.; Becker, D.F. (2015) Proline Metabolism Increases katG Expression and Oxidative Stress Resitance in Escherichia coli J Bacteriol 197(3): 431-40

Luo, M.; Christgen, S.; Sanyal, N.; Arentson, B.W.; Becker, D.F.; Tanner, J.J. (2014) Evidence that the C-Terminal Domain of a Type B PutA Protein Contributes to Aldehyde Dehydrogenase Activity and Substrate Channeling Biochemistry 53(35): 5661-73

Liang, X.; Dickman, M.B.; Becker, D.F. (2014) Proline Biosynthesis is Required for Endoplasmic Reticulum Stress Tolerance in Saccharomyces cerevisiae J Biol Chem 289(40): 27794-806

Arentson B.W.; Luo, M.; Pemberton T.A.; Tanner, J.J.; Becker, D.F. (2014) Kinetic and Structural Characterization of Tunnel-Perturbing Mutants in Bradyrhizobium japonicum Proline Utilization A Biochemistry 53(31): 5150-61

Singh, H.; Arentson, B.W.; Becker, D.F.; Tanner J.J. (2014) Structures of the PutA Peripheral Membrane Flavoenzyme Reveal a Dynamic Substrate-Channeling Tunnel and the Quinone-Binding Site PNAS 111(9): 3389-94

Pemberton T.A.; Srivastava, D.; Sanyal N.; Henzl, M.T.; Becker, D.F.; Tanner, J.J. (2014) Structural studies of yeast Δ(1)-pyrroline-5-carboxylate dehydrogenase (ALDH4A1): active site flexibility and oligomeric state. Biochemistry 53(8): 1350-9

Moxley, M.A.; Sanyal, N.; Krishnan, N.; Tanner, J.J.; Becker, D.F. (2014) Evidence for hysteretic substrate channeling in the proline dehydrogenase and Δ1-pyrroline-5-carboxylate dehydrogenase coupled reaction of proline utilization A (PutA) J Biol Chem 289(6) 3639-51

News and Alumni

Current Group Members
Joshua Floth selected to the ASBMB Honor Society for Undergraduate Researchers (April, 2016)
Shelbi Christgen awarded Milton E. Mohr Biotechnology Fellowship (May, 2015)
Joshua Floth awarded  Dr. Benjamin M. Sahagian Scholarship (May, 2015)
Shelbi Christgen awarded a MMoD Fellowship from the NIH T32 Program at UNL (2014-present)
Julia Zhang defended her PhD thesis and graduated in December 2015

Alumni
Ben Arentson (PhD, 2013) Quality Manager Poet Biorefining (ben.arentson@gmail.com)
Nikhilesh Sanyal (PhD, 2013) Global Data Science Analyst (nikhilesh1@gmail.com)
Recent Article by Nik (http://healthcare.globaldata.com/resources/expert-insights/pharmaceuticals/amid-suicide-concerns-a-new-astrazenecavaleant-deal-brings-brodalumab-back-to-life-as-a-promising-psoriasis-treatment)
Michael Moxley (PhD, 2012) Postdoctoral Associate, University of Michigan (mmox23@gmail.com)
Recent paper by Mike (http://www.ncbi.nlm.nih.gov/pubmed/26644471)
Sathish Natarajan (Postdoctoral Associate, 2009-2012) Assistant Professor, Department of Nutrition and Health Sciences,
University of Nebraska-Lincoln
Recent paper by Sathish (http://www.ncbi.nlm.nih.gov/pubmed/26610589)
Ashley Haile (MS, 2008) Nurse, University of Maryland Medical Center (ahaile1@umaryland.edu)

Navasona Krishnan (PhD, 2008)  Research Investigator, Cold Spring Harbor Laboratory (krishnn@cshl.edu)
News story on Nava's recent work (http://tbrnewsmedia.com/tonks-new-approach-to-rett-syndrome-at-cshl-shows-potential/)
Yuzhen Zhou (PhD, 2008) Professor, Huaiyin Normal University (zyz@hytc.edu.cn)
Weimin Zhang (Postdoctoral Associate, 2004-2007)
Manager, Analytical R&D, Impax Laboratories
Qi Cheng (Postdoctoral Associate, 2004-2006)
Srimevan Wanduragala (Postdoctoral Associate, 2003-2006) Senior Scientist-Analytical Development, Therapure Biopharma
Vinod Nair (Postdoctoral Associate, 2001-2004) Senior Electrochemist & Supercapacitor Expert, Calgon Carbon Corporation
(madhavanorama@gmail.com)
Weidong Zhu (PhD, 2005) Weidong Zhu <wdzhuwd@hotmail.com>
Berevan Baban (MS, 2003) Senior Research Tech, Washington University-St. Louis (bbaban@biochem.wustl.edu)
Recent paper from Berevan (http://www.ncbi.nlm.nih.gov/pubmed/25207746)
Padmanetra Bellur (MS, 2003) Biotechnology Professional