恒河猴晶状体核的蛋白质组学特征：与人类晶状体的相似性、年龄对蛋白质溶解度的影响以及翻译后修饰的趋势

Proteomic Characterization of the Rhesus Macaque Lens Nucleus: Similarity to Human Lens, Age Effects on Protein Solubility, and Trends in Post-Translational Modifications.

作者信息

Hayden Billy L, Kelley Owen, Zientek Keith, Reddy Ashok P, Wilmarth Phillip A, Munds Rachel, Montague Michael J, Martinez Melween I, Wollstein Gadi, Higham James P, Barron Arrambide Arturo O, Danias John, Melin Amanda D, David Larry L, Whitson Jeremy A

机构信息

Department of Biology, High Point University, High Point, North Carolina, United States.

Proteomics Shared Resource, Oregon Health & Science University, Portland, Oregon, United States.

出版信息

Invest Ophthalmol Vis Sci. 2025 Sep 2;66(12):28. doi: 10.1167/iovs.66.12.28.

DOI:10.1167/iovs.66.12.28

PMID:40938073

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12439503/

Abstract

PURPOSE

Proteomes of lens nuclei from young (4 years old) and old (15-16 years old) rhesus macaques (Macaca mulatta) were analyzed to determine similarity of the proteomic profile to that of human lenses, age-related differences in protein solubility, and association of various post-translational modifications with age and protein solubility.

METHODS

Lens core proteins were separated into water-soluble and water-insoluble fractions using aqueous buffer and centrifugation. The water-insoluble fraction was solubilized using sodium dodecyl sulfate (SDS). Proteins were processed using S-trap columns, and peptide digests were analyzed using high-resolution, label-free data-dependent acquisition (DDA) proteomics. Open modification searches were performed using MSFragger to identify possible post-translational modifications (PTMs). The number of modified peptide tandem mass spectra confidently assigned to samples by age or solubility were compared to find PTMs with statistically significant count differences.

RESULTS

The overall proteomic profile of rhesus macaque lenses was very similar to human lenses, consisting of 80.2% crystallins, 1.1% beaded filament proteins, and 18.7% other proteins. The crystallin fraction consisted of 27% alpha crystallins, 67.6% beta/gamma crystallins, and 5.4% taxon-specific psi crystallin. Glycolytic enzymes, beta/gamma crystallins, and a few glutathione-related enzymes were found to have age-related shifts to the water-insoluble fraction. There were significant differences in deamidation, dioxidation, carbamylation, carboxymethylation, and trioxidation based on age and/or solubility of proteins.

CONCLUSIONS

These data indicate a high level of conformity between rhesus macaque and human lens proteomes, and a few key differences. We identified several age-related differences in protein solubility and PTM that may contribute to lens pathology.

摘要

目的

分析幼年（4岁）和老年（15 - 16岁）恒河猴（猕猴）晶状体核的蛋白质组，以确定蛋白质组图谱与人类晶状体的相似性、蛋白质溶解度的年龄相关差异，以及各种翻译后修饰与年龄和蛋白质溶解度的关联。

方法

使用水性缓冲液和离心将晶状体核心蛋白分离为水溶性和水不溶性部分。水不溶性部分用十二烷基硫酸钠（SDS）溶解。蛋白质使用S-trap柱进行处理，肽段消化物使用高分辨率、无标记数据依赖采集（DDA）蛋白质组学进行分析。使用MSFragger进行开放修饰搜索以识别可能的翻译后修饰（PTM）。比较按年龄或溶解度可靠分配给样品的修饰肽串联质谱数量，以找到具有统计学显著计数差异的PTM。

结果

恒河猴晶状体的总体蛋白质组图谱与人类晶状体非常相似，由80.2%的晶状体蛋白、1.1%的珠状丝蛋白和18.7%的其他蛋白质组成。晶状体蛋白部分由27%的α晶状体蛋白、67.6%的β/γ晶状体蛋白和5.4%的分类群特异性ψ晶状体蛋白组成。发现糖酵解酶、β/γ晶状体蛋白和一些与谷胱甘肽相关的酶在年龄相关的情况下向水不溶性部分转移。基于蛋白质的年龄和/或溶解度，脱酰胺、二氧化、氨甲酰化、羧甲基化和三氧化存在显著差异。