Daneri-Castro Sergio N, Chandrasekar Balakumaran, Grosse-Holz Friederike M, van der Hoorn Renier A L, Roberts Thomas H
Faculty of Agriculture and Environment, Plant Breeding Institute, University of Sydney, Eveleigh, Australia.
The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, UK.
FEBS Lett. 2016 Sep;590(17):2956-62. doi: 10.1002/1873-3468.12320. Epub 2016 Aug 4.
During barley germination, the aleurone layer secretes most of the enzymes required to degrade the endosperm, many of which are yet to be characterized. We used activity-based protein profiling (ABPP) to detect a range of active enzymes extracted from aleurone layers isolated from grains of a commercial malting barley variety incubated with or without gibberellic acid (GA). Enzymes found to be induced by GA were putative aleurains, cathepsin-B-like proteases and serine hydrolases. By using an inhibitory sugar panel, a specific active retaining β-glycosidase in the barley aleurone was identified as a putative xylanase. Our results show that ABPP can be used rapidly to identify a variety of active enzyme isoforms in cereal aleurone without the need for enzyme purification.
在大麦发芽过程中，糊粉层分泌降解胚乳所需的大部分酶，其中许多酶尚未得到表征。我们使用基于活性的蛋白质谱分析（ABPP）来检测从商业麦芽大麦品种的籽粒中分离出的糊粉层中提取的一系列活性酶，这些籽粒分别在有或没有赤霉素（GA）的情况下进行培养。发现由GA诱导的酶是假定的 aleurains、组织蛋白酶B样蛋白酶和丝氨酸水解酶。通过使用抑制性糖组，在大麦糊粉层中一种具有特定活性的保留型β-糖苷酶被鉴定为假定的木聚糖酶。我们的结果表明，ABPP可用于快速鉴定谷物糊粉层中的多种活性酶同工型，而无需进行酶纯化。
