Identification and characterization of PaMTH1, a putative O-methyltransferase accumulating during senescence of Podospora anserina cultures
Identification and characterization of PaMTH1, a putative O-methyltransferase accumulating during senescence of Podospora anserina cultures
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DOI：
10.1007/s002940050520
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发表时间：
2000-03-01
期刊：
影响因子：
2.5
通讯作者：
Osiewacz, HD
中科院分区：
文献类型：
--
作者：
Averbeck, NB;Jensen, ON;Osiewacz, HD
差异蛋白质显示屏幕导致在标准条件下生长的Podospora anserina培养物的衰老过程中强烈积累的27-kDa蛋白质的鉴定。胰蛋白酶产生的片段的质谱分析的氨基酸序列的部分测定后，对简并引物推导和用于扩增编码的蛋白质的序列的部分。利用这些PCR产物从鹅绒委陵菜DNA文库中筛选特异性cDNA和基因组克隆。随后的DNA序列分析显示，27 kDa的蛋白质是由一个不连续的基因，PaMth 1，能够编码240个氨基酸。前三个氨基末端残基似乎是去除post-acetonally。推导的氨基酸序列显示出显着的同源性S-腺苷甲硫氨酸（SAM）依赖性甲基转移酶。我们推测，27 kDa的蛋白质，PaMTH 1，参与了与年龄相关的甲基化反应，保护老化的文化对增加氧化应激。
A differential protein display screen resulted in the identification of a 27-kDa protein which strongly accumulates during the senescence of Podospora anserina cultures grown under standard conditions. After partial determination of the amino-acid sequence by mass-spectrometry analysis of trypsin-generated fragments, pairs of degenerated primers were deduced and used to amplify parts of the sequence coding for the protein. These PCR products were utilized to select specific cDNA and genomic clones from DNA libraries of P. anserina. A subsequent DNA-sequence analysis revealed that the 27-kDa protein is encoded by a discontinuous gene, PaMth1, capable of coding for 240 amino acids. The first three amino-terminal residues appear to be removed post-translationally. The deduced amino-acid sequence shows significant homology to S-adenosylmethionine (SAM)-dependent methyltransferases. We hypothesize that the 27-kDa protein, PaMTH1, is involved in age-related methylation reactions protecting aging cultures against increasing oxidative stress.