Surmacz Liliana, Plochocka Danuta, Kania Magdalena, Danikiewicz Witold, Swiezewska Ewa
Biochim Biophys Acta. 2014 Feb;1841(2):240-50. doi: 10.1016/j.bbalip.2013.11.011.
cis-Prenyltransferases (CPTs) comprise numerous enzymes synthesizing isoprenoid hydrocarbon skeleton with isoprenoid units in the cis (Z) configuration. The chain-length specificity of a particular plant CPT is in most cases unknown despite thecomposition of the accumulated isoprenoids in the tissue of interest being well established. In this report AtCPT6, one of the nine Arabidopsis thaliana CPTs, is shown to catalyze the synthesis of a family of very short-chain polyisoprenoid alcohols of six, seven, and eight isoprenoid units, those of seven units dominating The product specificity of AtCPT6 was established in vivo following its expression in the heterologous system of the yeast Saccharomyces cerevisiae and was confirmed by the absence of specific products in AtCPT6 T-DNA insertion mutants and their overaccumulation in AtCPT6-overexpressing plants. These observations are additionally validated in silico using an AtCPT6 model obtained by homology modeling. AtCPT6 only partially complements the function of the yeast homologue of CPT-Rer2 since it restores the growth but not protein glycosylation in rer2delta yeast.This is the first in planta characterization of specific products of a plant CPT producing polyisoprenoids. Their distribution suggests that a joint activity of several CPTs is required to produce the complex mixture of polyisoprenoid alcohols found in Arabidopsis roots.
顺式异戊二烯转移酶(CPTs)包含众多能合成具有顺式(Z)构型异戊二烯单元的类异戊二烯烃骨架的酶。尽管目标组织中积累的类异戊二烯的组成已明确,但特定植物CPT的链长特异性在大多数情况下仍不清楚。在本报告中,拟南芥的九种CPTs之一AtCPT6被证明可催化合成由六个、七个和八个异戊二烯单元组成的非常短链的多异戊二烯醇家族,其中七个单元的多异戊二烯醇占主导。AtCPT6的产物特异性是在其在酿酒酵母异源系统中表达后在体内确定的,并通过AtCPT6 T-DNA插入突变体中特定产物的缺失及其在AtCPT6过表达植物中的过度积累得到证实。这些观察结果还通过使用通过同源建模获得的AtCPT6模型在计算机模拟中得到进一步验证。AtCPT6仅部分补充了CPT-Rer2酵母同源物的功能,因为它恢复了rer2delta酵母中的生长,但没有恢复蛋白质糖基化。这是首次在植物中对产生多异戊二烯类化合物的植物CPT的特定产物进行表征。它们的分布表明,需要几种CPTs的联合活性才能产生拟南芥根中发现的多异戊二烯醇的复杂混合物。