One-step site-specific S-alkylation of full-length caveolin-1: Lipidation modulates the topology of its C-terminal domain.
One-step site-specific S-alkylation of full-length caveolin-1: Lipidation modulates the topology of its C-terminal domain.
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DOI：
10.1002/pro.4791
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发表时间：
2023-11
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影响因子：
--
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Caveolin‐1是一种完整的膜蛋白，已知在转运到质膜时获得许多翻译后修饰。特别是，caveolin‐1在位于该蛋白C端结构域的三个半胱氨酸残基（C133、C143和C156）上被棕榈酰化，这可能具有结构和拓扑意义。本文描述了一种可靠的全长S‐烷基化小窝蛋白‐1的制备方法，它与体内观察到的棕榈酰化非常相似。HPLC和ESI‐LC‐MS分析验证了C16烷基基团被添加到含有1 （C133）、2 （C133和C143）和3 （C133、C143和C156）半胱氨酸残基的caveolin‐1结构中。结构体的圆二色光谱分析表明，S‐烷基化对蛋白质的整体螺旋度没有显著影响；然而，分子动力学模拟显示，在局部区域，螺旋度被S -烷基化正向或负向改变。此外，模拟结果表明，脂化抑制了C端结构域的拓扑混乱，导致双分子层内的分布以深度增加为特征。
Caveolin‐1 is an integral membrane protein that is known to acquire a number of posttranslational modifications upon trafficking to the plasma membrane. In particular, caveolin‐1 is palmitoylated at three cysteine residues (C133, C143, and C156) located within the C‐terminal domain of the protein which could have structural and topological implications. Herein, a reliable preparation of full‐length S‐alkylated caveolin‐1, which closely mimics the palmitoylation observed in vivo, is described. HPLC and ESI‐LC‐MS analyses verified the addition of the C16 alkyl groups to caveolin‐1 constructs containing one (C133), two (C133 and C143), and three (C133, C143, and C156) cysteine residues. Circular dichroism spectroscopy analysis of the constructs revealed that S‐alkylation does not significantly affect the global helicity of the protein; however, molecular dynamics simulations revealed that there were local regions where the helicity was altered positively or negatively by S‐alkylation. In addition, the simulations showed that lipidation tames the topological promiscuity of the C‐terminal domain, resulting in a disposition within the bilayer characterized by increased depth.
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影响因子：
3.7
作者：
Jo S;Kim T;Im W
通讯作者：
Im W
DOI：
10.1085/jgp.66.1.31
发表时间：
1975-07
期刊：
The Journal of general physiology
影响因子：
--
作者：
Mobley BA;Eisenberg BR
通讯作者：
Eisenberg BR
影响因子：
16.6
作者：
Hojo, Hironobu;Takei, Toshiki;Hirabayashi, Yoshio
通讯作者：
Hirabayashi, Yoshio
影响因子：
2.9
作者：
COMPTON, LA;JOHNSON, WC
通讯作者：
JOHNSON, WC
DOI：
10.1073/pnas.172196599
发表时间：
2002-08-20
影响因子：
11.1
作者：
Fernandez, I;Ying, YS;Anderson, RGW
通讯作者：
Anderson, RGW