BcrC from Bacillus subtilis acts as an undecaprenyl pyrophosphate phosphatase in bacitracin resistance
BcrC from Bacillus subtilis acts as an undecaprenyl pyrophosphate phosphatase in bacitracin resistance
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DOI：
10.1074/jbc.m413750200
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发表时间：
2005-08-12
影响因子：
4.8
通讯作者：
Denziot, F
中科院分区：
文献类型：
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作者：
Bernard, R;El Ghachi, M;Denziot, F
BCRC(Bs)蛋白，以前被称为YwoA，在大肠杆菌或枯草杆菌中的过表达使这些细菌能够耐受更高浓度的杆菌肽。推测BcrCBs是参与杆菌素耐药的三磷酸腺苷结合盒(ABC)转运体的跨膜结构域。然而，我们假设该蛋白具有十一碳烯基焦磷酸(UPP)磷酸酶活性，能够与杆菌肽竞争UPP。我们发现，重组His(6)-BCRC(Bs)蛋白在大肠杆菌中的过度表达(I)增加了细胞对杆菌肽的抵抗力，(Ii)使膜制剂中的UPP磷酸酶活性增加了600倍。我们溶解并制备了一种具有很强的UPP磷酸酶活性的电泳纯蛋白质。BCRC(Bs)属于2型磷脂酸磷酸酶(PAP2)超家族(PF01569)，与蛋白家族(Pfam)数据库中PF02673家族成员中已知的大肠杆菌Bac UPP磷酸酶完全不同。因此，BCRC(Bs)及其同源物在PAP2磷酸酶超家族中形成了一类新的蛋白质，并且它们可能都具有UPP磷酸酶活性。
Overexpression of the BcrC(Bs) protein, formerly called YwoA, in Escherichia coli or in Bacillus subtilis allows these bacteria to stand higher concentrations of bacitracin. It was suggested that BcrCBs was a membrane-spanning domain of an ATP binding cassette (ABC) transporter involved in bacitracin resistance. However, we hypothesized that this protein has an undecaprenyl pyrophosphate (UPP) phosphatase activity able to compete with bacitracin for UPP. We found that overexpression of a recombinant His(6)-BcrC(Bs) protein in E. coli (i) increased the resistance of the cells to bacitracin and (ii) increased UPP phosphatase activity in membrane preparations by 600-fold. We solubilized and prepared an electrophoretically pure protein exhibiting a strong UPP phosphatase activity. BcrC(Bs), which belongs to the type 2 phosphatidic acid phosphatase (PAP2) phosphatase superfamily (PF01569), differs totally from the already known BacA UPP phosphatase from E. coli, a member of the PF02673 family of the Protein family (Pfam) database. Thus, BcrC(Bs) and its orthologs form a new class of proteins within the PAP2 phosphatase superfamily, and likely all of them share a UPP phosphatase activity.