葵花叶片蛋白质组学分析样品制备方案评价。

Evaluation of sample preparation protocols for proteomic analysis of sunflower leaves.

机构信息

Group of Spectrometry, Sample Preparation and Mechanization (GEPAM), Institute of Chemistry, PO Box 6154, University of Campinas, UNICAMP, 13084-862, Campinas, SP, Brazil.

出版信息

Talanta. 2010 Feb 15;80(4):1545-51. doi: 10.1016/j.talanta.2009.06.016. Epub 2009 Jun 12.

DOI:10.1016/j.talanta.2009.06.016

PMID:20082813

Abstract

Six protocols for extraction of proteins from sunflower (Helianthus annuus L.) leaves were evaluated for their abilities in both removing interferents and attaining the best resolution in two-dimensional gel electrophoresis. "Classical" phenol extraction followed by precipitation with ammonium acetate in methanol displayed the most efficient protocol, which allowed the detection of 244 protein spots with ca. 485mug of protein in gel electrophoresis. Tandem mass spectrometry was performed to identify proteins in 61 spots, and cross species identification was used for this task. Proteins from twenty two spots were identified, and 12 of these proteins are up to now not included into the ExPASy sunflower protein databank.

摘要

六种从向日葵（Helianthus annuus L.）叶片中提取蛋白质的方法进行了评估，以评估它们在去除干扰物和在二维凝胶电泳中获得最佳分辨率方面的能力。“经典”的酚提取法，随后用甲醇中的醋酸铵沉淀，显示出最有效的方案，该方案允许在电泳中检测到约 244 个蛋白点，约 485μg 的蛋白。进行了串联质谱分析以鉴定 61 个斑点中的蛋白质，并使用交叉物种鉴定来完成这项任务。鉴定了 22 个斑点中的蛋白质，其中 12 种蛋白质迄今为止尚未包含在 ExPASy 向日葵蛋白质数据库中。