Identification of proteins from wild cardoon flowers (Cynara cardunculus L.) by a proteomic approach

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• 作者：Amal Ben Amira ; Julien Bauwens ; Edwin De Pauw…
• 关键词：Wild cardoon ; Protein extraction ; 2D gel electrophoresis ; Proteomic ; MALDI ; TOF MS ; Cardosin
• 刊名：Journal of Chemical Biology
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：10
• 期：1
• 页码：25-33
• 全文大小：
• 刊物主题：Physical Chemistry; Biological and Medical Physics, Biophysics; Cell Biology; Pharmacology/Toxicology; Biochemistry, general;
• 出版者：Springer Berlin Heidelberg
• ISSN：1864-6166
• 卷排序：10

文摘

Proteomic approach was applied to identify total proteins, particularly the enzymatic content, from wild cardoon flowers. As the selection of an appropriate sample preparation method is the key for getting reliable results, two different extraction/precipitation methods (trichloroacetic acid and phenol/ammonium acetate) were tested on fresh and lyophilized flowers. After two-dimensional electrophoresis (2D–E) separations, a better protein pattern was obtained after phenol extraction from lyophilized flowers. Only 46 % of the total analyzed spots resulted in a protein identification by mass spectrometry MALDI-TOF. Four proteases (cardosins A, E, G, and H), which have become a subject of great interest in dairy technology, were identified. They presented molecular weights and isoelectric points very close and high levels of homology between matched peptides sequences. The absence of the other cardosins (B, C, D, and F) could be an advantage, as it reduces the excessive proteolytic activity that causes bitter flavors and texture defects, during cheese making.