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两个重组的人类滑蛋白质之间的结构和功能比较:重组的人类蛋白质甘油-4 (rhPRG4) 与ECF84343之间的比较

Nikhil G Menon¹, Adam P Tanguay¹, Libo Zhou²

¹Biomedical Engineering Department, School of Dental Medicine, UConn Health, Farmington, CT, USA.

Experimental eye research

|September 7, 2023

概括

再组合的人类蛋白质甘4 (rhPRG4) 有效地治疗干眼疾病 (DED),与类似的蛋白质 (ECF843) 不一样. 在ECF843中的生物物理和功能差异解释了其降低的疗效,强调了蛋白质结构对治疗功能的重要性.

科学领域:

生物化学生物化学
生物物理学的生物物理.
眼科医生眼科眼科

背景情况:

蛋白质甘油4 (PRG4,滑素) 是用于眼皮表面滑和抗炎作用的关键糖蛋白.
重组人类PRG4 (rhPRG4) 在干眼疾病 (DED) 中显示出临床疗效.
不同的复合形式ECF843在临床试验中与载体相比,没有显著改善DED.

研究的目的:

在生物物理和功能上比较rhPRG4和ECF843.
了解rhPRG4和ECF843在DED中的不同临床疗效的结构基础.
为了阐明PRG4变体的结构功能关系.

主要方法:

动态光散射 (DLS) 和水力动态直径和电荷的泽塔潜力.
用多角度光散射 (SEC-MALS) 进行尺寸排除色谱,以测量尺寸和分子量.
拉曼光谱法用于键结构和分析超离心法 (AUC) 用于沉积.
功能性测试包括MMP-9抑制,边界滑,原1结合,酸盐晶体抑制和细胞粘附.

主要成果:

ECF843的水力动力直径较小,负电荷也比 rhPRG4.4 低.
与rhPRG4.4相比,SEC-MALS发现ECF843具有更小和更少的多元器件.
拉曼光谱显示,ECF843具有更多的二硫化物键,而AUC显示它更紧.
在功能上,ECF843在MMP-9抑制,滑,原1结合,尿酸盐晶体抑制和细胞粘附预防方面效果较差.

结论:

在rhPRG4和ECF843.4之间存在显著的结构和功能差异.
这些分子差异可能解释了在DED试验中观察到的不同临床结果.
蛋白质结构对于眼睛表面应用中PRG4的治疗功能至关重要.

关键词:

欧洲银行基金84343 卢布里辛是一种乳制品.蛋白质甘油4 (PRG4) 是一种蛋白质甘油.

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