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http://dx.doi.org/10.1007/s11816-010-0150-3

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)
Publication Information
Plant Biotechnology Reports / v.4, no.4, 2010 , pp. 329-334 More about this Journal
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
Agrobacterium sp. strain CP4; 5-Enolpyruvylshikimate-3-phosphate synthase (EPSPS); Glyphosate; Herbicide resistance; Proline residue;
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1 Kang K, Kim M (2006a) Expression of the glyphosate resistant gene, cp4-epsps, through plastid transformation in rice (Oryza sativa L.). J Plant Biotechnol 33:75-84   DOI   ScienceOn
2 Bull HB, Breese K (1974) Surface tension of amino acid solutions: a hydrophobicity scale of the amino acid residues. Arch Biochem Biophys 161:665-670   DOI   ScienceOn
3 Duke SO, Powles SB (2008) Glyphosate: a once-in-a-century herbicide. Pest Manag Sci 64:319-325   DOI   ScienceOn
4 Eisenberg D, McLachlan AD (1986) Solvation energy in protein folding and binding. Nature 319:199-203   DOI   ScienceOn
5 Kang K, Kim M (2006b) Single amino acid changes in the 5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4 and Synechocystis sp. PCC6803 alter enzyme activity and glyphosate sensitivity. Agric Chem Biotechnol 49:65-68
6 Balbach J, Schmid FX (2000) Proline isomerization and its catalysis in protein folding. In: Pain RH (ed) Mechanisms of protein folding, 2nd edn. Oxford University Press, UK
7 Padgette SR, Re DB, Barry GF, Eichholtz DE, Delannay X, Fuchs RL, Kishore GM, Fraley RT (1996) New weed control opportunities: development of soybeans with a Roundup Ready gene. In: Duck SO (ed) Herbicide-resistant crops: agricultural, economic, environmental, regulatory and technological aspects. CRC Press, Boca Raton, USA, pp 53-84
8 Park H, Hilsenbeck JL, Kim HJ, Shuttleworth WA, Park YH, Evans JN, Kang K (2003) Structural studies of Streptococcus pneumoniae EPSP synthase in unliganded state, tetrahedral intermediatebound state and S3P-GLP-bound state. Mol Microbiol 51:963-971
9 Schulz GE, Schirmer RH (1979) Principle of protein structure. Springer, New York
10 Steinrucken HC, Amerhein N (1984) 5-Enolpyruvylshikimate-3- phosphate synthase of Klebsiella pneumonia 2. Inhibition by glyphosate [N-(phosphonomethyl)glycine]. Eur J Biochem 143:351-357   DOI   ScienceOn
11 Kishore GM, Shah DM (1988) Amino acid biosynthesis inhibitors as herbicides. Annu Rev Biochem 57:627-663   DOI   ScienceOn
12 Lanzetta PA, Alvarez LJ, Reinach PS, Candia OA (1979) An improved assay for nanomole amounts of inorganic phosphate. Anal Biochem 100:95-97   DOI   ScienceOn
13 Herrmann KM (1995) The shikimate pathway: early steps in the biosynthesis of aromatic compounds. Plant Cell 7:907-919
14 Malik J, Barry G, Kishore G (1989) The herbicide glyphosate. Biofactors 2:17-25
15 Eschenburg SM, Healy ML, Priestman MA, Lushington GH, Schonbrunn E (2002) How the mutation glycine96 to alanine confers glyphosate insensitivity to 5-enolpyruvyl shikimate-3- phosphate synthase from Escherichia coli. Planta 216:129-135   DOI   ScienceOn
16 Garcia-Effron G, Katiyar SK, Park S, Edlind TD, Perlin DS (2008) A naturally occurring proline-to-alanine amino acid change in Fks1p in Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis accounts for reduced echinocandin susceptibility. Antimicrob Agents Chemother 52:2305-2312   DOI   ScienceOn
17 Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR (1989) Sitedirected mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51-59   DOI   ScienceOn
18 Funke T, Han H, Healy-Fried ML, Fisher M, Schonbrunn E (2006) Molecular basis for the herbicide resistance of Roundup Ready crops. Proc Natl Acad Sci USA 103:13010–13015   DOI   ScienceOn
19 Schonbrunn E, Eschenburg S, Shuttleworth WA, Schloss JV, Amrhein N, Evans JN, Kabsch W (2001) Interaction of the herbicide glyphosate with its target enzyme 5-enolpyruvylshikikate 3-phosphate synthase in atomic detail. Proc Natl Acad Sci USA 98:1376-1380   DOI   ScienceOn
20 Boocock MR, Coggins JR (1983) Kinetics of 5-enolpyruvylshikimate- 3-phosphate synthase inhibition by glyphosate. FEBS Lett 154:127-133   DOI   ScienceOn
21 Kelly RF, Richards FM (1987) Replacement of proline-76 with alanine eliminates the slowest kinetic phase in thioredoxin folding. Biochem 26:6765-6774   DOI   ScienceOn