Brian Mooney

Research Assistant Professor of Biochemistry
Associate Director, Proteomics Center

Email:	mooneyb@missouri.edu
Phone:	(573) 884-7374
Fax:	(573) 884-9676
Office:	214 Bond Life Sciences Center
Mailing Address:	Proteomics Center 214 LSC, 1201 E. Rollins St. University of Missouri-Columbia Columbia, MO 65211
Research Areas:	Proteomics of protein expression and assembly; metabolic engineering.
Webpage:	http://proteomics.missouri.edu

Educational Background

Research Description

Production of Biodegradable Plastics in Plants - The polyhydroxyalkanoate (PHA) copolymer, P(3HB-co-3HV), is a type of biodegradable plastic demonstrated to be useful in the manufacture of bottles, bags, disposable razors, flatware, etc. Additionally, medical uses for PHA, e.g. implants, drug delivery, have been proposed. To date there has been limited success in using plants as a means of PHA production by targeting the bacterial enzymes responsible for PHA synthesis to the chloroplasts of Arabidopsis thaliana. This resulted in the production of low levels (3% dry wt.) of the PHA copolymer in the chloroplasts. One of the precursors for the PHA co-polymer, acetyl-CoA, is present naturally in the chloroplasts at sufficient levels to support PHA production. However, the other precursor, propionyl-CoA, was produced at a very low efficiency by the endogenous plastid pyruvate dehydrogenase complex (PDC). The branched-chain α-ketoacid dehydrogenase enzyme (BCE1) is capable of production of propionyl-CoA at a much higher efficiency that the plastid PDC. However the BCE1 enzyme is located in a different compartment of the plant cell, the mitochondria.

The objective of the project (being conducted in Douglas D. Randall's lab by Elizabeth Hoyos) is to alter the intra-cellular location of the Arabidopsis thaliana BCE1 (AtBCE1) enzyme from mitochondria to plastids. We have generated plastid-targeted AtBCE1 by replacing the mitochondrial-targeting portions of the two proteins that make up the AtBCE1 (AtBCE1α and AtBCE1β) with plastid-targeting sequences derived from the A. thaliana small subunit of RuBisCO. In addition, we have replaced the region of AtBCE1β responsible for binding to the central core enzyme, BCE2, with a region capable of binding plastid PDC E2. We have confirmed plastid-targeting by western blot of chloroplasts isolated from transgenic plants. We have also confirmed (although indirectly through enzyme assays) the ability of the plastid-targeted AtBCE1 to form a heterotetrameric complex as well as its ability to bind to the endogenous plastid PDC E2. Once we have confirmed stable, heritable, and functional, plastid-targeted AtBCE1 we will cross these plants with those that posses the (bacterial) genes for biodegradable plastic synthesis. We will then asses levels of biodegradable plastic produced in these transgenic plants.

Selected Publications

Mooney B.P., Miernyk J.A., and Randall D.D. (2002) The complex fate of alpha-ketoacids. In: Annu. Rev. Plant Biol. 2002, D.P. Delmer, H.J. Bohnert, S Merchant, eds, Vol. 53: 357-375

Mooney B.P., Krishnan H.B., and Thelen J.J. (2004) High-throughput peptide mass fingerprinting of soybean seed proteins: automated workflow and utility of UniGene expressed sequence tag databases for protein identification. Phytochemistry 65:1733-44

Wan J, Torres M, Ganapathy A, Thelen J, DaGue BB, Mooney B, Xu D, Stacey G (2005) Proteomic analysis of soybean root hairs after infection by Bradyrhizobium japonicum. Mol Plant Microbe Interact 18: 458-67

Mooney B.P., Miernyk J.A., Greenlief C.M., Thelen, J.J. (2006) Using quantitative proteomics of Arabidopsis roots and leaves to predict metabolic activity. Physiologia Plantarum 128: 237-250.

McCaw DL, Chan AS, Stegner AL, Mooney B, Bryan JN, Turnquist SE, Henry CJ, Alexander H, and Alexander S (2007) Proteomics of Canine Lymphoma Identifies Potential Cancer-Specific Protein Markers. Clin Cancer Res 13:2496-2503