| Cloning and functional expression of the small subunit of acetolactate synthase from Nicotiana plumbaginifolia. | |
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MedLine Citation:
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PMID: 10487214 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Acetolactate synthase (ALS) is the first committed step of branched-chain amino acid biosynthesis in plants and bacteria. The bacterial holoenzyme has been well characterized and is a tetramer of two identical large subunits (LSUs) of 60 kDa and two identical small subunits (SSUs) ranging in molecular mass from 9 to 17 kDa depending on the isozyme. The enzyme from plants is much less well characterized. Attempts to purify the protein have yielded an enzyme which appears to be an oligomer of LSUs, with the potential existence of a SSU for the plant enzyme remaining a matter of considerable speculation. We report here the discovery of a cDNA clone that encodes a SSU of plant ALS based upon the homology of the encoded peptide with various bacterial ALS SSUs. The plant ALS SSU is more than twice as large as any of its prokaryotic homologues and contains two domains that each encode a full-length copy of the prokaryotic SSU polypeptide. The cDNA clone was used to express Nicotiana plumbaginifolia SSU in Escherichia coli. Mixing a partially purified preparation of this SSU with the LSU of ALS from either N. plumbaginifolia or Arabidopsis thaliana results in both increased specific activity and increased stability of the enzymic activity. These results are consistent with those observed for the bacterial enzyme in similar experiments and represent the first functional demonstration of the existence of a SSU for plant ALS. |
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Authors:
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H P Hershey; L J Schwartz; J P Gale; L M Abell |
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Publication Detail:
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Type: Comparative Study; Journal Article |
Journal Detail:
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Title: Plant molecular biology Volume: 40 ISSN: 0167-4412 ISO Abbreviation: Plant Mol. Biol. Publication Date: 1999 Jul |
Date Detail:
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Created Date: 1999-10-04 Completed Date: 1999-10-04 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9106343 Medline TA: Plant Mol Biol Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 795-806 Citation Subset: IM |
Affiliation:
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Agricultural Products Department, E.I. du Pont de Nemours, Inc., Stine-Haskell Research Laboratory, Newark, DE 19714, USA. |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/AJ234901 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acetolactate Synthase
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chemistry,
genetics*,
metabolism Amino Acid Sequence Base Sequence Cloning, Molecular Escherichia coli / enzymology Genetic Vectors Isoenzymes / chemistry, genetics, metabolism Macromolecular Substances Molecular Sequence Data Molecular Weight Plants, Toxic* Recombinant Fusion Proteins / biosynthesis, chemistry, metabolism Seeds / enzymology Sequence Alignment Sequence Homology, Amino Acid Tobacco / enzymology*, genetics* |
| Chemical | |
Reg. No./Substance:
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0/Isoenzymes; 0/Macromolecular Substances; 0/Recombinant Fusion Proteins; EC 2.2.1.6/Acetolactate Synthase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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