Identification and characterization of a serine protease from wheat leaves
详细信息 查看全文
- 作者：Tao Fan ; Natalia V. Bykova ; Christof Rampitsch…
- 关键词：Subtilisin ; like protease ; Programmed cell death ; Stress ; Wheat
- 刊名：European Journal of Plant Pathology
- 出版年：2016
- 出版时间：October 2016
- 年：2016
- 卷：146
- 期：2
- 页码：293-304
- 全文大小：1,050 KB
- 刊物类别：Biomedical and Life Sciences
- 刊物主题：Life Sciences
Plant Pathology
Plant Sciences
Ecology - 出版者：Springer Netherlands
- ISSN：1573-8469
- 卷排序：146
文摘
A putative serine protease with a potential role in the plant biotic and abiotic stress response was purified from wheat leaf apoplastic fluid and partially characterized. Following two-dimensional electrophoresis a protein of Mr = 75 k and a pI of 4.2 to 4.5 was observed. This protein displayed in-gel protease activity and was specifically inhibited by phenylmethanesulfonyl fluoride and partially inhibited by Ca2+ and Zn2+, but not by E-64 or leupeptin. An internal tryptic fragment of 13 amino acids was identified by MALDI QqTOF MS/MS, and this peptide showed a high level of homology (85–100 % identity) to a highly conserved region of known plant subtilisin-like proteases. We demonstrated that the protease activity increased until a late stage of wheat leaf development and increased in response to heat shock. In both cases Rubisco large subunit was degraded with time. Protease activity was also increased during biotic stress. Leaves challenged with leaf rust (Puccinia triticina), showed an approximately three fold increase in protease activity during an incompatible interaction, compared to activity in mock-inoculated leaves and to leaves in a compatible leaf rust interaction. These results suggest that the expression of this serine protease could be involved in the defense response against both abiotic and biotic stresses.