Lipase secretion by bacterial hybrid ATP-binding cassette exporters: Molecular recognition of the LipBCD, PrtDEF, and HasDEF exporters
Lipase secretion by bacterial hybrid ATP-binding cassette exporters: Molecular recognition of the LipBCD, PrtDEF, and HasDEF exporters
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DOI：
10.1128/jb.179.15.4754-4760.1997
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发表时间：
1997-08-01
影响因子：
3.2
通讯作者：
Omori, K
中科院分区：
文献类型：
--
作者：
Akatsuka, H;Binet, R;Omori, K
粘质沙雷氏菌分泌几种蛋白质，如脂肪酶利帕、金属蛋白酶PrtA和血红素结合蛋白HasA，其是血红素获取所需的，通过两个N-末端信号肽独立系统，其被分类为细菌ATP结合盒（ABC）输出者。一个是HasA的ABC输出者，由ABC蛋白HasD、膜融合蛋白（MFP）HasE、第二种由LipB（ABC蛋白）、LipC（MFP）和LipD（OMP）组成，在大肠杆菌重组克隆中促进利帕和PrtA的分泌。大肠杆菌细胞携带的E.在该细菌中存在不同的系统以及这些系统的各种底物，允许研究异源Has、Lip和Prt系统以及Has-Lip和Lip-Prt杂交输出者在真实宿主以及E.为此目的，从S. coli中分离出lipB-、lipC-和lipD-缺陷突变体。marcescens 8000及其分泌的利帕和PrtA进行了分析。这表明，一个独特的出口商，唇器，在S。marcescens分泌利帕和PrtA。检测了杂交输出子的HasA和利帕分泌。LipB-HasE-HasF输出子允许利帕分泌，但不允许HasA分泌，表明ABC蛋白LipB负责底物特异性，利帕、HasA和E。通过杂交输出蛋白的分泌分析表明，在这些系统中，MFP和OMP之间存在特异性相互作用。这些遗传实验表明，ABC蛋白和MFP之间的特异性相互作用是形成活性输出蛋白所必需的。
Serrartia marcescens secretes several proteins, such as the lipase LipA, the metalloprotease PrtA, and the heme-binding protein HasA, which is required for heme acquisition, through two N-terminal signal peptide independent systems that are classified as bacterial ATP-binding cassette (ABC) exporters, One is the ABC exporter for HasA, consisting of the ABC protein HasD, the membrane fusion protein (MFP) HasE, and the outer membrane protein (OMP) HasF, The second, composed of LipB (an ABC protein), LipC (an MFP), and LipD (an OMP), promotes secretion of LipA and PrtA in Escherichia coli recombinant clones, PrtA, which shows homology to the Erwinia chrysanthemi metalloproteases, is efficiently secreted by E. coli cells carrying the E. chrysanthemi ABC exporter PrtD (ABC protein)-PrtE (MFP)-PrtF (OMP), The existence of distinct systems in this bacterium and of various substrates for these systems allowed the study of protein secretion by heterologous Has, Lip, and Prt systems and by Has-Lip and Lip-Prt hybrid exporters in the genuine host as well as in E. coli, For that purpose, lipB-, lipC-, and lipD-deficient mutants were isolated from S. marcescens 8000 and their secretion of LipA and PrtA was analyzed. This demonstrated that a unique exporter, the Lip apparatus, in S. marcescens secretes both LipA and PrtA. Hybrid exporters were tested for secretion of HasA and LipA, The LipB-HasE-HasF exporter allowed secretion of LipA but not HasA, showing that the ABC protein LipB is responsible for the substrate specificity, LipA, HasA, and E. chrysanthemi PrtC were secreted via heterologous exporters and via some hybrid exporters, Analysis of secretion via hybrid exporters showed that specific interactions occur between MFPs and OMPs in these systems, These genetic experiments demonstrated that specific interactions between the ABC protein and the MFP are required for the formation of active exporters.