Penicillin-binding protein 2x of Streptococcus pneumoniae: the mutation Ala707Asp within the C-terminal PASTA2 domain leads to destabilization.

Penicillin-binding protein 2x of Streptococcus pneumoniae: the mutation Ala707Asp within the C-terminal PASTA2 domain leads to destabilization.
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肺炎链球菌青霉素结合蛋白 2x:C 端 PASTA2 结构域内的 Ala707Asp 突变导致不稳定

DOI:
10.1089/mdr.2014.0082
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发表时间:
2014
影响因子:
2.6
通讯作者:
D. Denapaite
D. Denapaite
中科院分区:
医学4区
文献类型:
--
作者:
Schweizer;K. Peters;C. Stahlmann;R. Hakenbeck;D. Denapaite

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肺炎链球菌青霉素结合蛋白2x (PBP2x)是一种参与肽聚糖组装最后阶段的酶,对细菌的生长和生存至关重要。PBP2x定位于分裂位点,这一过程依赖于它的青霉素结合蛋白和丝氨酸-苏氨酸激酶相关(PASTA)结构域,之前通过GFP-PBP2x在活细胞中证实了这一点。在这项研究中,分离出一株突变菌株,其中GFP-PBP2x融合蛋白不定位于分裂位点,并且含有全长GFP-PBP2x的减少量。我们现在证明这种缺陷是由于PBP2x c端PASTA2结构域内的点突变引起的。突变蛋白在活细胞中的β -内酰胺结合、功能和定位方面进行了详细分析。我们证明该突变严重影响gfp标记的PBP2x变体,并使其对蛋白酶/伴侣蛋白HtrA敏感。
Streptococcus pneumoniaepenicillin-binding protein 2x (PBP2x) is an enzyme involved in the last stages of peptidoglycan assembly and essential for bacterial growth and survival. PBP2x localizes to the division site, a process that depends on itsPenicillin-Binding ProteinAndSerine-Threonine-kinaseAssociated (PASTA) domains, which was previously demonstrated via GFP-PBP2x in living cells. During this study a mutant strain was isolated in which the GFP-PBP2x fusion protein did not localize at division sites and it contained reduced amounts of the full-length GFP-PBP2x. We now show that this defect is due to a point mutation within the C-terminal PASTA2 domain of PBP2x. The mutant protein was analyzed in detail in terms of beta-lactam binding, functionality, and localization in live cells. We demonstrate that the mutation affects the GFP-tagged PBP2x variant severely and renders it susceptible to the protease/chaperone HtrA.
DOI: 10.1111/j.1365-2958.1996.tb02556.x
发表时间: 1996
影响因子: 3.6
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