Complex formation between recombinant ATP sulfurylase and APS reductase of Allium cepa (L.)
- 作者：Mathew Cumming ; Susanna Leung ; John McCallum ; Michael T. McManus
- 关键词：Adenosine-5′ ; -phosphosulfate reductase ; ATP sulfurylase ; Protein– ; protein interactions ; Sulfate assimilation ; Sulfur nutrition ; Allium cepa (L.)
- 刊名：FEBS Letters
- 出版年：2007
- 期刊代码：70_00145793
- 类别：bio
- 出版时间：4 September 2007
- 卷：581
- 期：22
- 页码：4139-4147
- 文件大小：358 K
摘要
Recombinant ATP sulfurylase (AcATPS1) and adenosine-5′-phosphosulfate reductase (AcAPR1) from Allium cepa have been used to determine if these enzymes form protein–protein complexes in vitro. Using a solid phase binding assay, AcAPR1 was shown to interact with AcATPS1. The AcAPR1 enzyme was also expressed in E. coli as the N-terminal reductase domain (AcAPR1-N) and the C-terminal glutaredoxin domain (AcAPR1-C), but neither of these truncated proteins interacted with AcATPS1. The solid-phase interactions were confirmed by immune-precipitation, where anti-AcATPS1 IgG precipitated the full-length AcAPR1 protein, but not AcAPR1-N and AcAPR1-C. Finally, using the ligand binding assay, full-length AcATPS1 has been shown to bind to membrane-localised full-length AcAPR1. The significance of an interaction between chloroplastidic ATPS and APR in A. cepa is evaluated with respect to the control of the reductive assimilation of sulfate.