Document Detail


Isotope labeling of RuBisCO subunits provides in vivo information on subcellular biosynthesis and exchange of amino acids between compartments.
MedLine Citation:
PMID:  22292468     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
The architecture of plant metabolism includes substantial duplication of metabolite pools and enzyme catalyzed reactions in different subcellular compartments. This poses challenges for understanding the regulation of metabolism particularly in primary metabolism and amino acid biosynthesis. To explore the extent to which amino acids are made in single compartments and to gain insight into the metabolic precursors from which they derive, we used steady state (13) C labeling and analyzed labeling in protein amino acids from plastid and cytosol. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is a major component of green tissues and its large and small subunits are synthesized from different pools of amino acids in the plastid and cytosol respectively. Developing Brassica napus embryos were cultured in the presence of [U-(13) C]-sucrose, [U-(13) C]-glucose, [U-(13) C]-glutamine, or [U-(13) C]-alanine to generate proteins. The large (LSU) and small (SSU) RuBisCO subunits were isolated and the labeling in their constituent amino acids was analyzed by GC mass spectrometry. Amino acids including alanine, glycine and serine exhibited different (13) C enrichment in the LSU and SSU, demonstrating that these pools have different metabolic origins and are not isotopically equilibrated between the plastid and cytosol on the time scale of cellular growth. Potential extensions of this novel approach to other macromolecules, organelles and cell types of eukaryotes are discussed. Published 2012. This article is a U.S. Government work and is in the public domain in the USA.
Authors:
Doug K Allen; Russell W Laclair; John B Ohlrogge; Yair Shachar-Hill
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-1-31
Journal Detail:
Title:  Plant, cell & environment     Volume:  -     ISSN:  1365-3040     ISO Abbreviation:  -     Publication Date:  2012 Jan 
Date Detail:
Created Date:  2012-2-1     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9309004     Medline TA:  Plant Cell Environ     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Published 2012. This article is a U.S. Government work and is in the public domain in the USA.
Affiliation:
USDA-ARS, Plant Genetics Research Unit, St Louis MO 63132 (D.K.A.); Donald Danforth Plant Science Center, St Louis MO 63132 (D.K.A.); Great Lakes Bioenergy Research Center (R.W.L., J.B.O., Y.S.H.) and Department of Plant Biology, Michigan State University, East Lansing, MI 48824 (R.W.L., J.B.O., Y.S.H.).
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