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相关概念视频

Export of Misfolded Proteins out of the ER01:32

Export of Misfolded Proteins out of the ER

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After folding, the ER assesses the quality of secretory and membrane proteins. The correctly folded proteins are cleared by the calnexin cycle for transport to their final destination, while misfolded proteins are held back in the ER lumen. The ER chaperones attempt to unfold and refold the misfolded proteins but sometimes fail to achieve the correct native conformation. Such terminally misfolded proteins are then exported to the cytosol by ER-associated degradation or ERAD pathway for...
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相关实验视频

Updated: May 15, 2025

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
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Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

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蛋白酶的高通量活动重编程 (HARP)

Samantha G Martinusen, Ethan W Slaton, Seyednima Ajayebi

    bioRxiv : the preprint server for biology
    |April 8, 2025
    PubMed
    概括

    高通量蛋白酶活动重编程 (HARP) 平台有效地发现了强大的蛋白酶抑制剂. 这种以酵母为基础的屏隔离了抑制性宏分子,使新药候选药物的快速识别成为可能.

    科学领域:

    • 生物化学 生物化学
    • 分子生物学分子生物学
    • 药物发现 药物发现 药物发现

    背景情况:

    • 开发特定的蛋白酶抑制剂是具有挑战性的,因为需要广泛的表征.
    • 现有的平台优先考虑高亲和度结合剂,往往缺少罕见的,强大的抑制剂.

    研究的目的:

    • 开发一个新的平台,有效地发现蛋白酶抑制剂.
    • 使用功能屏幕对特定蛋白酶的低纳米抑制剂进行隔离.

    主要方法:

    • 开发了蛋白酶高通量活动重编程 (HARP) 平台,一种基于酵母的功能屏幕.
    • 与可选择细胞表面表型的蛋白酶抑制相结合,用于高通量选.
    • 利用结构建模和深度测序进行抑制剂分析.

    主要成果:

    • 成功分离低纳米阻碍纳米体,对抗烟草蚀刻病毒蛋白酶 (TEVp) 和人类卡利克林 6.
    • 确定了一种罕见的7.6 nM Ki非竞争性TEVp抑制剂.
    • 在抑制剂发现中,HARP展示了高动态范围和分辨率.

    结论:

    • 哈普 (HARP) 是一个主要的平台,用于发现对酶标的调节性宏分子.

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    High Throughput Quantitative Expression Screening and Purification Applied to Recombinant Disulfide-rich Venom Proteins Produced in E. coli
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  • 该平台有助于分离强效和选择性蛋白酶抑制剂.
  • 提供了关于抑制剂有效性的分子决定因素的见解.