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使用温和提取方法鉴定哺乳动物视网膜中的纤毛蛋白

Identifying Ciliary Proteins in Mammalian Retinas using a Gentle Extraction Method.

作者信息

Fredrick Adeline S, Claussen Erin R, Fischer Samantha N, Balasanyants Samson, Rajaraman Akshaya, Rosner Andi C, Drew Kevin

机构信息

Biological Sciences, University of Illinois at Chicago, Chicago, Illinois, United States.

出版信息

MicroPubl Biol. 2024 Jun 13;2024. doi: 10.17912/micropub.biology.001218. eCollection 2024.

Abstract

Mutations in retinal primary cilia are responsible for human blindness but the mechanisms are not fully understood (Wheway et al., 2014). Characterizing the proteome of an organelle such as cilia, is a fruitful way to understand its function but methods often require considerable sample quantities. Here we develop a method to isolate the primary cilia of photoreceptor cells from bovine retinas. Through LC/MS/MS proteomics analysis we identify proteins enriched for cilia function including ciliopathy disease. This study shows our method can be used to isolate retinal primary cilia to obtain sufficient quantities of native protein samples.

摘要

视网膜初级纤毛的突变是导致人类失明的原因,但其中的机制尚未完全明确(Wheway等人,2014年)。对诸如纤毛这样的细胞器的蛋白质组进行表征,是了解其功能的有效途径,但这些方法通常需要大量样本。在此,我们开发了一种从牛视网膜中分离光感受器细胞初级纤毛的方法。通过液相色谱/串联质谱(LC/MS/MS)蛋白质组学分析,我们鉴定出了富含纤毛功能(包括纤毛病相关)的蛋白质。这项研究表明,我们的方法可用于分离视网膜初级纤毛,以获得足够数量的天然蛋白质样本。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dc06/11211918/0082b2be98c4/25789430-2024-micropub.biology.001218.jpg

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