Human defensin 5 disulfide array mutants: disulfide bond deletion attenuates antibacterial activity against Staphylococcus aureus.
Human defensin 5 disulfide array mutants: disulfide bond deletion attenuates antibacterial activity against Staphylococcus aureus.
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DOI：
10.1021/bi201043j
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发表时间：
2011-09-20
期刊：
影响因子：
2.9
通讯作者：
Nolan, Elizabeth M.
中科院分区：
文献类型：
--
作者：
Wanniarachchi, Yoshitha A.;Kaczmarek, Piotr;Wan, Andrea;Nolan, Elizabeth M.
人α-防御素5（HD 5，HD 5 ox）是一种富含半胱氨酸的宿主防御肽，由小肠潘氏细胞表达和释放，对多种革兰氏阴性和革兰氏阳性细菌菌株具有抗菌活性。为了确定其二硫键排列对结构、抗微生物活性和蛋白水解稳定性的贡献，在大肠杆菌中过量表达了一系列HD 5双突变肽，其中对应于天然二硫键的半胱氨酸残基对（Cys 3-Cys 31，Cys 5-Cys 20，Cys 10-Cys 30）突变为Ser或Ala残基。大肠杆菌，纯化和表征。还评价了六个突变肽HD 5 [Serhexa]，其中所有六个天然Cys残基均被Ser残基取代。去除单个天然S-S键会影响氧化折叠和区域异构化、抗菌活性、革兰氏阴性菌膜透化和蛋白水解稳定性。然而，大多数HD 5突变肽显示出低微摩尔的抗革兰氏阴性E.大肠杆菌ATCC 25922中的菌落计数试验中，野生型二硫键阵列对于针对革兰氏阳性S.金黄色葡萄球菌ATCC 25923。单个二硫键的去除减弱了观察到的HD 5 ox针对该革兰氏阳性细菌菌株的活性。这一观察结果支持了这样的观点，即HD 5 ox的抗菌作用机制对于革兰氏阴性和革兰氏阳性物种是不同的（Wei，G.; de Leeuw，E.，Pazgier，M.，袁伟，Zou，G.，王杰，Ericksen，B.，Lu，W.- Y的;莱勒河一、Lu，W.（2009）J.Biol.Chem.284，29180-29192），并且天然二硫键阵列是其针对S.金黄色。
Human α-defensin 5 (HD5, HD5ox to specify the oxidized and disulfide linked form) is a 32-residue cysteine-rich host-defense peptide, expressed and released by small intestinal Paneth cells, that exhibits antibacterial activity against a number of Gram-negative and –positive bacterial strains. To ascertain the contributions of its disulfide array to structure, antimicrobial activity, and proteolytic stability, a series of HD5 double mutant peptides where pairs of cysteine residues corresponding to native disulfide linkages (Cys3—Cys31, Cys5—Cys20, Cys10—Cys30) were mutated to Ser or Ala residues were overexpressed in E. coli, purified and characterized. A hexa mutant peptide, HD5[Serhexa], where all six native Cys residues are replaced by Ser residues was also evaluated. Removal of a single native S—S linkage influences oxidative folding and regioisomerization, antibacterial activity, Gram-negative bacterial membrane permeabilization, and proteolytic stability. Whereas the majority of the HD5 mutant peptides show low-micromolar activity against Gram-negative E. coli ATCC 25922 in colony counting assays, the wild-type disulfide array is essential for low-micromolar activity against Gram-positive S. aureus ATCC 25923. Removal of a single disulfide bond attenuates the activity observed for HD5ox against this Gram-positive bacterial strain. This observation supports the notion that the HD5ox mechanism of antibacterial action differs for Gram-negative and Gram-positive species (Wei, G.; de Leeuw, E., Pazgier, M., Yuan, W., Zou, G., Wang, J., Ericksen, B., Lu, W.-Y.; Lehrer, R. I.; Lu, W. (2009) J. Biol. Chem. 284, 29180-29192), and that the native disulfide array is a requirement for its activity against S. aureus.
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影响因子：
2.9
作者：
Fázio, MA;Oliveira, VX;Miranda, A
通讯作者：
Miranda, A
DOI：
10.1007/978-1-60761-594-1_4
发表时间：
2010-01-01
期刊：
ANTIMICROBIAL PEPTIDES: METHODS AND PROTOCOLS
影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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