Mapping the Anopheles gambiae odorant binding protein 1 (AgamOBP1) using modeling techniques, site directed mutagenesis, circular dichroism and ligand binding assays.
Mapping the Anopheles gambiae odorant binding protein 1 (AgamOBP1) using modeling techniques, site directed mutagenesis, circular dichroism and ligand binding assays.
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DOI：
10.1016/j.bbapap.2012.04.011
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发表时间：
2012-08
影响因子：
3.2
通讯作者：
Eliopoulos, E.
中科院分区：
文献类型：
--
作者：
Rusconi, B.;Maranhao, A. C.;Fuhrer, J. P.;Krotee, P.;Choi, S. H.;Grun, F.;Thireou, T.;Dimitratos, S. D.;Woods, D. F.;Marinotti, O.;Walter, M. F.;Eliopoulos, E.
关键词：
撒哈拉以南非洲的主要疟疾病媒是冈比亚按蚊。该物种是疟疾控制措施的主要目标。蚊子主要通过嗅觉找到人类，但与宿主寻找行为相关的分子机制仍然很大程度上未知。为了进一步了解A.冈比亚气味结合蛋白1（AgamOBP 1），我们结合在电脑蛋白质结构建模和定点诱变，以产生16个AgamOBP 1蛋白类似物含有单点突变的兴趣。圆二色性（CD）和配体结合试验提供了必要的数据来探测点突变对配体结合和AgamOBP 1的整体结构的影响。突变的AgamOBP 1变体的远紫外CD光谱显示出有序α-螺旋结构的显著降低（高达22%）和无序α-螺旋结构的增加（高达15%），无规卷曲（无序）结构的变化极小。在突变Y 54 A、Y122 A和W114 Q中，从结合位点去除芳香族侧链显著降低了N-苯基-1-萘胺结合。几个非芳香族突变（L15 T、L19 T、L58 T、L58 Y、M84 Q、M84 K、H111 A、Y122 A和L124 T）引起蛋白质构象的变化，随后影响配体结合。该研究为AgamOBP 1折叠和配体结合特性中特定氨基酸的计算机预测功能提供了经验证据。
The major malaria vector in Sub-Saharan Africa is the Anopheles gambiae mosquito. This species is a key target of malaria control measures. Mosquitoes find humans primarily through olfaction, yet the molecular mechanisms associated with host-seeking behavior remain largely unknown. To further understand the functionality of A. gambiae odorant binding protein 1 (AgamOBP1), we combined in silico protein structure modeling and site-directed mutagenesis to generate 16 AgamOBP1 protein analogues containing single point mutations of interest. Circular dichroism (CD) and ligand-binding assays provided data necessary to probe the effects of the point mutations on ligand binding and the overall structure of AgamOBP1. Far-UV CD spectra of mutated AgamOBP1 variants displayed both substantial decreases to ordered α-helix structure (up to 22%) and increases to disordered α-helix structure(up to 15%) with only minimal changes in random coil (unordered) structure. In mutations Y54A, Y122A and W114Q, aromatic side chain removal from the binding site significantly reduced N-phenyl-1-naphthylamine binding. Several non-aromatic mutations (L15T, L19T, L58T, L58Y, M84Q, M84K, H111A, Y122A and L124T) elicited changes to protein conformation with subsequent effects on ligand binding. This study provides empirical evidence for the in silico predicted functions of specific amino acids in AgamOBP1 folding and ligand binding characteristics.
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影响因子：
4.8
作者：
Lartigue, A;Gruez, A;Cambillau, C
通讯作者：
Cambillau, C
影响因子：
1.6
作者：
Thireou, Trias;Atlamazoglou, Vassilios;Eliopoulos, Elias
通讯作者：
Eliopoulos, Elias
影响因子：
8
作者：
Tsitsanou, K. E.;Thireou, T.;Zographos, S. E.
通讯作者：
Zographos, S. E.
DOI：
10.1002/prot.1155
发表时间：
2001-12-01
期刊：
PROTEINS-STRUCTURE FUNCTION AND GENETICS
影响因子：
--
作者：
Berezovsky, IN;Kirzhner, VM;Trifonov, EN
通讯作者：
Trifonov, EN
影响因子：
2.9
作者：
PROVENCHER, SW;GLOCKNER, J
通讯作者：
GLOCKNER, J