In this project we use stable isotope labeling of intact plant tissues to analyze the pathways of fatty acid and lipid biosynthesis from precursors such as [13C] or [13C, 18O]acetate, [18O]water and [13C]carbon dioxide. These experiments give us information on the precursor pools, fluxes through precursor pools, and the relevance of putative hydrolytic reactions in lipid biosynthesis. Already, preliminary data for leaves suggests that the bulk acetate pool is not the primary carbon source for fatty acid synthesis and that long-chain fatty acyl export from the plastid does indeed occur via a hydrolytic mechanism. We have used MS to check the isotope composition of precursor and identification of novel natural products). More importantly, we have identified several mass spectrometry experiments which will help us unlock the fundamental question of the number of fatty acid and lipid synthesizing systems operating in the cell of the developing oilseed. These experiments will require development of mass spectrometry methods, and in particular (1) the measurement of molecular species of neutral lipids in INDIVIDUAL oil bodies, and (2) cryo-sectioning of seed tissue and mapping the 2-D distribution of lipid molecular species at as high a resolution as possible (preferably approaching resolution of a single cell), using MALDI-TOF or other focusable soft ionization technique. Both these types of experiment would expand the applicability of mass spectroscopy for the study of lipid biosynthesis and require an exciting and interactive collaboration with the MSU Mass Spectrometry Facility.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
3P41RR000480-28S1
Application #
6258848
Study Section
Project Start
1997-06-01
Project End
1999-11-30
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
28
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Michigan State University
Department
Type
DUNS #
193247145
City
East Lansing
State
MI
Country
United States
Zip Code
48824
Peri, S P; Bhadti, V S; Somerville-Armstrong, K S et al. (1999) Affinity reagents for cross-linking hemoglobin: bis(phenoxycarbonylethyl)phosphinic acid (BPCEP) and bis(3-nitrophenoxycarbonylethyl)phosphinic acid (BNCEP). Hemoglobin 23:1-20
Chen, H M; Sood, R; Hosmane, R S (1999) An efficient, short synthesis and potent anti-hepatitis B viral activity of a novel ring-expanded purine nucleoside analogue containing a 5:7-fused, planar, aromatic, imidazo[4,5-e][1,3]diazepine ring system. Nucleosides Nucleotides 18:331-5
Bretner, M; Beckett, T D; Sood, R K et al. (1999) Substrate/inhibition studies of bacteriophage T7 RNA polymerase with the 5'-triphosphate derivative of a ring-expanded ('fat') nucleoside possessing potent antiviral and anticancer activities. Bioorg Med Chem 7:2931-6
Agasimundin, Y S; Mumper, M W; Hosmane, R S (1998) Inhibitors of glycogen phosphorylase b: synthesis, biochemical screening, and molecular modeling studies of novel analogues of hydantocidin. Bioorg Med Chem 6:911-23
Hosmane, R S; Peri, S P; Bhadti, V S et al. (1998) Bis[2-(4-carboxyphenoxy)carbonylethyl]phosphinic acid (BCCEP): a novel affinity reagent for the beta-cleft modification of human hemoglobin. Bioorg Med Chem 6:767-83
Rajappan, V P; Hosmane, R S (1998) Analogues of azepinomycin as inhibitors of guanase. Nucleosides Nucleotides 17:1141-51
Hosmane, R S; Hong, M (1997) How important is the N-3 sugar moiety in the tight-binding interaction of coformycin with adenosine deaminase? Biochem Biophys Res Commun 236:88-93
Lopez-Lara, I M; Orgambide, G; Dazzo, F B et al. (1993) Characterization and symbiotic importance of acidic extracellular polysaccharides of Rhizobium sp. strain GRH2 isolated from acacia nodules. J Bacteriol 175:2826-32
Watson, J T; Kayganich, K (1989) Novel sample preparation for analysis by electron capture negative ionization mass spectrometry. Biochem Soc Trans 17:254-7
Kassel, D B; Kayganich, K A; Watson, J T et al. (1988) Utility of ion source pretreatment with chlorine-containing compounds for enhanced performance in gas chromatography/negative ionization mass spectrometry. Anal Chem 60:911-7

Showing the most recent 10 out of 11 publications