Plant Biotechnology Reports

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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The three proline residues (P25, P242, and P434) of Agrobacterium CP4 5-enolpyruvylshikimate-3-phosphate synthase are crucial for the enzyme activity

  • Kang, Kyung-Su (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Jin, Yong-Mei (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Jeon, Hye-Sung (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Park, Sang-Ryoung (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Research Institute for Agriculture and Life Sciences, Seoul National University) ;
  • Song, Dae-Geun (Natural Products Research Center, Korea Institute of Science and Technology Gangneung Institute) ;
  • Lee, Joo-Young (Natural Products Research Center, Korea Institute of Science and Technology Gangneung Institute) ;
  • Pan, Cheol-Ho (Natural Products Research Center, Korea Institute of Science and Technology Gangneung Institute) ;
  • Kim, Min-Kyun (Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, Research Institute for Agriculture and Life Sciences, Seoul National University)
  • Received : 2010.08.11
  • Accepted : 2010.08.29
  • Published : 2010.12.30
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Abstract

Multiple sequence alignments showed that the prolines at the 25th, 129th, 153rd, 242nd, 322nd, and 434th amino acids in 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium sp. strain CP4 are strongly conserved in various prokaryotic EPSPS proteins. Single point mutations of the conserved prolines to alanine (P25A, P153A, P242A, P322A, and P434A) were introduced in the CP4 EPSPS in order to investigate the importance of the conserved prolines for the enzyme properties. The point mutations caused decreases in substrate binding affinity and catalytic efficiency as well as the glyphosate resistance, in general. Especially, the 25th and 242nd prolines located in the polypeptide hinges connecting top and bottom domains of CP4 EPSPS as well as the 434th proline at the C-terminus of the enzyme turned out to be crucial for the enzyme activity.

Keywords

References

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