Diener A C, Li H, Zhou W, Whoriskey W J, Nes W D, Fink G R
Whitehead Institute for Biomedical Research, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02142, USA.
Plant Cell. 2000 Jun;12(6):853-70. doi: 10.1105/tpc.12.6.853.
The side chain in plant sterols can have either a methyl or ethyl addition at carbon 24 that is absent in cholesterol. The ethyl addition is the product of two sequential methyl additions. Arabidopsis contains three genes-sterol methyltransferase 1 (SMT1), SMT2, and SMT3-homologous to yeast ERG6, which is known to encode an S-adenosylmethionine-dependent C-24 SMT that catalyzes a single methyl addition. The SMT1 polypeptide is the most similar of these Arabidopsis homologs to yeast Erg6p. Moreover, expression of Arabidopsis SMT1 in erg6 restores SMT activity to the yeast mutant. The smt1 plants have pleiotropic defects: poor growth and fertility, sensitivity of the root to calcium, and a loss of proper embryo morphogenesis. smt1 has an altered sterol content: it accumulates cholesterol and has less C-24 alkylated sterols content. Escherichia coli extracts, obtained from a strain expressing the Arabidopsis SMT1 protein, can perform both the methyl and ethyl additions to appropriate sterol substrates, although with different kinetics. The fact that smt1 null mutants still produce alkylated sterols and that SMT1 can catalyze both alkylation steps shows that there is considerable overlap in the substrate specificity of enzymes in sterol biosynthesis. The availability of the SMT1 gene and mutant should permit the manipulation of phytosterol composition, which will help elucidate the role of sterols in animal nutrition.
植物甾醇中的侧链在碳24位上可以有甲基或乙基加成,而胆固醇中没有这种加成。乙基加成是两个连续甲基加成的产物。拟南芥含有三个与酵母ERG6同源的基因——甾醇甲基转移酶1(SMT1)、SMT2和SMT3,已知酵母ERG6编码一种依赖S-腺苷甲硫氨酸的C-24甾醇甲基转移酶,催化一次甲基加成。SMT1多肽是拟南芥这些同源物中与酵母Erg6p最相似的。此外,在erg6中表达拟南芥SMT1可使酵母突变体恢复SMT活性。smt1植株有多种缺陷:生长和育性差、根对钙敏感以及胚胎形态发生异常。smt1的甾醇含量发生了改变:它积累了胆固醇,C-24烷基化甾醇含量减少。从表达拟南芥SMT1蛋白的菌株中获得的大肠杆菌提取物,可以对合适的甾醇底物进行甲基和乙基加成,尽管动力学不同。smt1缺失突变体仍能产生烷基化甾醇,且SMT1能催化两个烷基化步骤,这一事实表明甾醇生物合成中酶的底物特异性存在相当大的重叠。SMT1基因和突变体的可用性应有助于操纵植物甾醇的组成,这将有助于阐明甾醇在动物营养中的作用。