Del Buono Daniele, Prinsi Bhakti, Espen Luca, Scarponi Luciano
Dipartimento di Scienze Agrarie e Ambientali, Universita degli Studi di Perugia, Borgo XX Giugno 72, 06121 Perugia, Italy.
J Agric Food Chem. 2009 Sep 9;57(17):7924-30. doi: 10.1021/jf901681q.
The effect of treatments with four herbicides and a safener on the activity of triosephosphate isomerase (TPI) extracted from shoots of Italian ryegrass was investigated. It was found that atrazine and fluorodifen, herbicides which interfere with photosynthesis, caused a decrease in measured enzyme activity. In addition, the in vitro effect of oxidized glutathione (GSSG), a compound produced in situations of oxidative stress, on TPI activity was investigated. It was shown that GSSG was a strong inhibitor of enzyme activity, at low concentrations in a dose-time-dependent manner. The enzyme extracts were submitted to chromatographic purifications and to two-dimensional electrophoresis. Some spots had molecular masses ranging between 20 and 30 kDa and were characterized and identified by LC-ESI-MS/MS as TPIs. The mass spectrometry also made it possible to identify the presence of cysteine residues that could be subjected to S-glutathionylation, which regulate the enzyme activity.
研究了四种除草剂和一种安全剂处理对从意大利黑麦草芽中提取的磷酸丙糖异构酶（TPI）活性的影响。发现干扰光合作用的除草剂阿特拉津和氟乐灵会导致测得的酶活性降低。此外，还研究了氧化型谷胱甘肽（GSSG）（一种在氧化应激情况下产生的化合物）对TPI活性的体外作用。结果表明，GSSG是酶活性的强抑制剂，在低浓度下呈剂量-时间依赖性。酶提取物经过色谱纯化和二维电泳。一些斑点的分子量在20至30 kDa之间，通过LC-ESI-MS/MS表征并鉴定为TPI。质谱分析还能够鉴定出可能发生S-谷胱甘肽化修饰的半胱氨酸残基的存在，这种修饰可调节酶的活性。
