Borrelia burgdorferi HtrA: evidence for twofold proteolysis of outer membrane protein p66.
Borrelia burgdorferi HtrA: evidence for twofold proteolysis of outer membrane protein p66.
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DOI：
10.1111/mmi.13221
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发表时间：
2016-01
影响因子：
3.6
通讯作者：
Benach JL
中科院分区：
文献类型：
--
作者：
Coleman JL;Toledo A;Benach JL
在原核生物中，高温要求A(HtrA)丝氨酸蛋白酶家族的成员在周质中发挥作用，降解受损或不正确折叠的膜蛋白。伯氏疏螺旋体是莱姆病的病原体，编码单一的HtrA同源物。对Burgdorferi B31A3和一株高表达HtrA的菌株(A3HtrAOE)的双向电泳分析发现，A3HtrAOE中有一种下调蛋白，其质量、等电点和MALDI-TOF谱与外膜蛋白P66一致。细胞裂解物中的p66和htrA被分成耐洗涤剂的膜，膜中含有被称为脂筏的富含胆固醇的糖脂膜区域，这表明htrA和p66可能也一起存在于脂筏中。这与我们实验室以前的工作一致，该工作表明HtrA和P66是伯氏杆菌外膜囊泡的成分。HtrA在体外降解p66，A3HtrAOE在体内表达降低水平的p66。荧光共聚焦显微镜显示HtrA和p66共定位于细胞膜。HtrA和p66的结合证实了它们可以有效地相互作用，它们的蛋白酶/底物关系为这种相互作用提供了功能上的相关性。与野生型B31A3相比，A3HtrAOE还显示p66转录水平降低，这表明HtrA对p66的调节可能发生在多个水平。
In prokaryotes, members of the High Temperature Requirement A (HtrA) family of serine proteases function in the periplasm to degrade damaged or improperly folded membrane proteins. Borrelia burgdorferi, the agent of Lyme disease, codes for a single HtrA homolog. Two-dimensional electrophoresis analysis of B. burgdorferi B31A3 and a strain that over-expresses HtrA (A3HtrAOE) identified a down-regulated protein in A3HtrAOE with a mass, pI and MALDI-TOF spectrum consistent with outer membrane protein p66. P66 and HtrA from cellular lysates partitioned into detergent-resistant membranes, which contain cholesterol-glycolipid-rich membrane regions known as lipid rafts, suggesting that HtrA and p66 may reside together in lipid rafts also. This agrees with previous work from our laboratory, which showed that HtrA and p66 are constituents of B. burgdorferi outer membrane vesicles. HtrA degraded p66 in vitro and A3HtrAOE expressed reduced levels of p66 in vivo. Fluorescence confocal microscopy revealed that HtrA and p66 co-localize in the membrane. The association of HtrA and p66 establishes that they could interact efficiently and their protease/substrate relationship provides functional relevance to this interaction. A3HtrAOE also showed reduced levels of p66 transcript in comparison to wild-type B31A3, indicating that HtrA-mediated regulation of p66 may occur at multiple levels.
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影响因子：
6.7
作者：
Crowley JT;Toledo AM;LaRocca TJ;Coleman JL;London E;Benach JL
通讯作者：
Benach JL
影响因子：
3.4
作者：
Barcena-Uribarri, Ivan;Thein, Marcus;Benz, Roland
通讯作者：
Benz, Roland
影响因子：
3.2
作者：
Bunikis, J;Luke, CJ;Barbour, AG
通讯作者：
Barbour, AG
影响因子：
3.1
作者：
Defoe, G;Coburn, J
通讯作者：
Coburn, J
影响因子：
3.1
作者：
COLEMAN, JL;ROGERS, RC;BENACH, JL
通讯作者：
BENACH, JL