Native chemical ligation at Asx-Cys, Glx-Cys: chemical synthesis and high-resolution X-ray structure of ShK toxin by racemic protein crystallography.
Native chemical ligation at Asx-Cys, Glx-Cys: chemical synthesis and high-resolution X-ray structure of ShK toxin by racemic protein crystallography.
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DOI：
10.1021/ja4046795
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发表时间：
2013-08-14
影响因子：
15
通讯作者：
Kent SB
中科院分区：
文献类型：
--
作者：
Dang B;Kubota T;Mandal K;Bezanilla F;Kent SB
我们重新研究了−Gln/Glu-Cys− [Glx-Cys]和−Asn/Asp-Cys− [Asx-Cys]位点天然化学连接的效用。使用改进的硫代芳基催化剂4-巯基苯乙酸（MPAA），可以在−Gln-Cys−和Asn-Cys−位点进行天然化学连接而没有副反应。在优化之后，在−Glu-Cys−处的连接也可以用作连接位点，具有最小水平的副产物形成。然而，-Asp-Cys-不适合用作天然化学连接的位点，因为会形成大量的β-连接副产物。在−Gln-Cys−上进行天然化学连接的可行性使ShK毒素蛋白分子的对映体形式的收敛全化学合成成为可能。D-ShK蛋白分子在阻断Kv1.3通道方面的活性比L-ShK蛋白分子低约50，000倍。外消旋蛋白质晶体学用于获得高分辨率的X-射线衍射数据的ShK毒素。结构通过直接方法解决，并显示出显着的差异，从以前报道的NMR结构中的ShK蛋白分子的某些区域。
We have re-examined the utility of native chemical ligation at −Gln/Glu-Cys− [Glx-Cys] and −Asn/Asp-Cys− [Asx-Cys] sites. Using the improved thioaryl catalyst 4-mercaptophenylacetic acid (MPAA), native chemical ligation could be performed at −Gln-Cys− and Asn-Cys− sites without side reactions. After optimization, ligation at a −Glu-Cys− could also be used as a ligation site, with minimal levels of byproduct formation. However, −Asp-Cys− is not appropriate for use as a site for native chemical ligation because of formation of significant amounts of β-linked byproduct. The feasibility of native chemical ligation at −Gln-Cys− enabled a convergent total chemical synthesis of the enantiomeric forms of the ShK toxin protein molecule. The D-ShK protein molecule was ~50,000-fold less active in blocking the Kv1.3 channel than the L-ShK protein molecule. Racemic protein crystallography was used to obtain high resolution X-ray diffraction data for ShK toxin. The structure was solved by direct methods and showed significant differences from the previously reported NMR structures in some regions of the ShK protein molecule.
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影响因子：
15
作者：
Pentelute BL;Gates ZP;Tereshko V;Dashnau JL;Vanderkooi JM;Kossiakoff AA;Kent SB
通讯作者：
Kent SB
影响因子：
56.9
作者：
DAWSON, PE;MUIR, TW;KENT, SBH
通讯作者：
KENT, SBH
影响因子：
14.9
作者：
Maiti, R;Van Domselaar, GH;Wishart, DS
通讯作者：
Wishart, DS
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
15
作者：
Mandal K;Pentelute BL;Tereshko V;Kossiakoff AA;Kent SB
通讯作者：
Kent SB