O-GlcNAc转移酶在四重复 (TPR) 超螺旋中使用阿斯巴拉金梯子识别蛋白质基质
Zebulon G Levine1, Chenguang Fan1, Michael S Melicher1
1Department of Microbiology and Immunobiology , Harvard Medical School , Boston , Massachusetts 02115 , United States.
Journal of the American Chemical Society
|February 28, 2018
概括
酶O-GlcNAc转移酶 (OGT) 在其TPR域中使用阿斯巴拉金梯子来识别和糖化蛋白质基质. 这一发现澄清了OGT如何选择O-GlcNAcylation的目标.
科学领域:
- 生物化学
- 分子生物学
- 葡萄糖生物学
背景情况:
- 酶O-GlcNAc转移酶 (OGT) 修改了许多核和细胞质蛋白.
- 由于缺乏共识序列,OGT的基质识别机制在很大程度上是未知的.
研究的目的:
- 研究OGT如何识别其多种蛋白质基质.
- 确定阿斯巴拉金梯子在基质选择中的四基重复 (TPR) 域中的作用.
主要方法:
- 开发一种化学酶蛋白微阵列测定,以评估数千种蛋白质的OGT活性.
- 在OGT TPR域内对5个黄素残留物的位点定向突变发生.
主要成果:
- 突变阿斯巴拉金梯子显著降低了OGT在微阵列上糖化蛋白质基质的能力.
- 细胞提取物中的蛋白质基质的O- GlcNAcylation也明显减弱.
- OGT保留了对短的活性,这表明梯子对蛋白质识别至关重要.
结论:
- 在OGT TPR域内,阿斯巴拉金梯子对于识别其大多数蛋白质基质至关重要.
- 这种相互作用发生在接近催化域的TPR光线中.
- 该研究阐明了OGT基质特异性的关键机制.
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