Short N-terminal sequences package proteins into bacterial microcompartments
Short N-terminal sequences package proteins into bacterial microcompartments
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DOI：
10.1073/pnas.0913199107
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发表时间：
2010-04-20
影响因子：
11.1
通讯作者：
Bobik, Thomas A.
中科院分区：
文献类型：
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作者：
Fan, Chenguang;Cheng, Shouqiang;Bobik, Thomas A.
数百种细菌产生蛋白质微区室（MCP），其通过限制具有毒性或挥发性中间体的代谢途径的酶而充当简单的细胞器。关于细菌MCP的一个基本的未回答的问题是酶如何包装在形成其外表面的蛋白质壳内。在这里，我们报告说，一个短的N-末端肽是必要的和足够的包装酶进入MCP的内腔参与B-1,2-依赖的1，2-丙二醇利用（Pdu MCP）。从通常在Pdu MCP内发现的丙醛脱氢酶（PduP）的N末端缺失10或14个氨基酸，显著损害包装，对其酶活性的影响最小。PduP的18个N-末端氨基酸与GFP、GST或麦芽糖结合蛋白的融合导致它们在MCP内的包封。生物信息学分析揭示了两个额外的Pdu蛋白和来自两个不相关的MCP的三个蛋白的N-末端延伸，表明N-末端肽可用于将蛋白质包装成不同的MCP。MCP组装原理在自然界和生物技术中的潜在用途进行了讨论。
Hundreds of bacterial species produce proteinaceous microcompartments (MCPs) that act as simple organelles by confining the enzymes of metabolic pathways that have toxic or volatile intermediates. A fundamental unanswered question about bacterial MCPs is how enzymes are packaged within the protein shell that forms their outer surface. Here, we report that a short N-terminal peptide is necessary and sufficient for packaging enzymes into the lumen of an MCP involved in B-12-dependent 1,2-propanediol utilization (Pdu MCP). Deletion of 10 or 14 amino acids from the N terminus of the propionaldehyde dehydrogenase (PduP) enzyme, which is normally found within the Pdu MCP, substantially impaired packaging, with minimal effects on its enzymatic activity. Fusion of the 18 N-terminal amino acids from PduP to GFP, GST, or maltose-binding protein resulted in their encapsulation within MCPs. Bioinformatic analyses revealed N-terminal extensions in two additional Pdu proteins and three proteins from two unrelated MCPs, suggesting that N-terminal peptides may be used to package proteins into diverse MCPs. The potential uses of MCP assembly principles in nature and in biotechnology are discussed.