Functional plasticity of paralogous diterpene synthases involved in conifer defense

被引:143
作者
Keeling, Christopher I. [1 ]
Weisshaar, Sabrina [1 ]
Lin, Roy P. C. [1 ]
Bohlmann, Joerg [1 ]
机构
[1] Univ British Columbia, Michael Smith Lab, Vancouver, BC V6T 1Z4, Canada
关键词
biochemical evolution; natural product biosynthesis; plant defense; terpene diversity; site-directed mutagenesis;
D O I
10.1073/pnas.0709466105
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The diversity of terpenoid compounds produced by plants plays an important role in mediating various plant-herbivore, plant-pollinator, and plant-pathogen interactions. This diversity has resulted from gene duplication and neofunctionalization of the enzymes that synthesize and subsequently modify terpenes. Two diterpene synthases in Norway spruce (Picea abies), isopimara,diene synthase and levopimaradiene/abietadiene synthase, provide the hydrocarbon precursors for most of the diterpene resin acids found in the defensive oleoresin of conifers. Although these paralogous enzymes are 91% identical at the amino acid level, one is a single-product enzyme, whereas the other is a multiproduct enzyme that forms completely different products. We used a rational approach of homology modeling, protein sequence comparison, domain swapping, and a series of reciprocal site-directed mutagenesis to identify the specific residues that direct the different product outcomes. A one-amino acid mutation switched the levopimaradiene/abietadiene synthase into producing isopimaradiene and sandaracopimaradiene and none of its normal products. Four mutations were sufficient to reciprocally reverse the product profiles for both of these paralogous enzymes while maintaining catalytic efficiencies similar to the wild-type enzymes. This study illustrates how neofunctionalization can result from relatively minor changes in protein sequence, increasing the diversity of secondary metabolites important for conifer defense.
引用
收藏
页码:1085 / 1090
页数:6
相关论文
共 34 条
[1]  
ADAMS R. P., 2007, IDENTIFICATION ESSEN
[2]   Plant terpenoid synthases: Molecular biology and phylogenetic analysis [J].
Bohlmann, J ;
Meyer-Gauen, G ;
Croteau, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (08) :4126-4133
[3]   Structural biology and chemistry of the terpenoid cyclases [J].
Christianson, David W. .
CHEMICAL REVIEWS, 2006, 106 (08) :3412-3442
[4]   DITERPENOID RESIN ACID BIOSYNTHESIS IN CONIFERS - CHARACTERIZATION OF 2 CYTOCHROME P450-DEPENDENT MONOOXYGENASES AND AN ALDEHYDE DEHYDROGENASE INVOLVED IN ABIETIC ACID BIOSYNTHESIS [J].
FUNK, C ;
CROTEAU, R .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1994, 308 (01) :258-266
[5]   Identifying and manipulating structural determinates linking catalytic specificities in terpene synthases [J].
Greenhagen, Bryan T. ;
O'Maille, Paul E. ;
Noel, Joseph P. ;
Chappell, Joe .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (26) :9826-9831
[6]   SWISS-MODEL and the Swiss-PdbViewer: An environment for comparative protein modeling [J].
Guex, N ;
Peitsch, MC .
ELECTROPHORESIS, 1997, 18 (15) :2714-2723
[7]   An EXCEL template for calculation of enzyme kinetic parameters by non-linear regression [J].
Hernandez, A ;
Ruiz, MT .
BIOINFORMATICS, 1998, 14 (02) :227-228
[8]   Mutational analysis of a monoterpene synthase reaction:: Altered catalysis through directed mutagenesis of (-)-pinene synthase from Abies grandis [J].
Hyatt, DC ;
Croteau, R .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2005, 439 (02) :222-233
[9]   Rational conversion of substrate and product specificity in a Salvia monoterpene synthase:: Structural insights into the evolution of terpene synthase function [J].
Kampranis, Sotirios C. ;
Ioannidis, Daphne ;
Purvis, Alan ;
Mahrez, Walid ;
Ninga, Ederina ;
Katerelos, Nikolaos A. ;
Anssour, Samir ;
Dunwell, Jim M. ;
Degenhardt, Joerg ;
Makris, Antonios M. ;
Goodenough, Peter W. ;
Johnson, Christopher B. .
PLANT CELL, 2007, 19 (06) :1994-2005
[10]   Diterpene resin acids in conifers [J].
Keeling, Christopher I. ;
Bohlmann, Joerg .
PHYTOCHEMISTRY, 2006, 67 (22) :2415-2423