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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Characterization of Two Forms of Acetolactate Synthase from Barley

  • Yoon, Jong-Mo (School of Life Sciences and Biotechnology Research Institute, Chungbuk National University) ;
  • Yoon, Moon-Young (Department of Chemistry, Hanyang University) ;
  • Kim, Young-Tae (Department of Microbiology, Pukyong National University) ;
  • Choi, Jung-Do (School of Life Sciences and Biotechnology Research Institute, Chungbuk National University)
  • Received : 2003.02.24
  • Accepted : 2003.03.13
  • Published : 2003.09.30
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Abstract

Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of valine, leucine, and isoleucine. ALS is the target site for several classes of herbicides, including sulfonylureas, imidazolinones, and triazolopyrimidines. Two forms of ALS (designated ALS I and ALS II) were separated from barley shoots by heparin affinity column chromatography. The molecular masses of native ALS I and ALS II were determined to be 248 kDa and 238 kDa by nondenaturing gel electrophoresis and activity staining. Similar molecular masses of two forms of ALS were confirmed by a Western blot analysis. SDS-PAGE and Western blot analysis showed that the molecular masses of the ALS I and ALS II subunits were identical - 65 kDa. The two ALS forms exhibited different properties with respect to the values of $K_m$, pI and optimum pH, and sensitivity to inhibition by herbicides sulfonylurea and imidazolinone as well as to the feedback regulation by the end-product amino acids Val, Leu, and Ile. These results, therefore, suggest that the two ALS forms are not different polymeric forms of the same enzyme, but isozymes.

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References

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