Heterogeneity in proline hydroxylation of fibrillar collagens observed by mass spectrometry.
Heterogeneity in proline hydroxylation of fibrillar collagens observed by mass spectrometry.
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DOI：
10.1371/journal.pone.0250544
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Xu Y
中科院分区：
文献类型：
--
作者：
Kirchner M;Deng H;Xu Y
胶原蛋白是细胞外基质中的主要蛋白质，在组织发育和功能中起着重要作用。胶原蛋白也是其生物合成中加工最多的蛋白质之一。胶原蛋白最显著的翻译后修饰（PTM）是胶原蛋白三螺旋的特征性（Gly-Xaa-Yaa）重复氨基酸序列的Y-位置中Pro残基的羟基化。最近使用质谱法（MS）和串联MS测序（MS/MS）的研究揭示了X-位置（X-Hyp）中Pro残基的意外羟基化。新鉴定的X-Hyp残基似乎在位置和占据百分比上高度异质。为了了解新的X-Hyps的动态性质及其对MS和MS/MS在胶原蛋白研究中的应用的潜在影响，我们使用标准MS和MS/MS技术对四种不同的胶原蛋白样品进行了采样。我们发现来自不同生物体和/或组织的相同胶原蛋白的PTM程度存在相当大的差异。大鼠尾腱I型胶原蛋白在X-位置的Pro的过度羟基化和Y-位置的Pro的羟基化不足方面特别可变。相比之下，在来自人胎盘的I型和III型胶原中仅观察到少数意外的PTM。一些观察结果在相同样品的不同测序工作之间是不可重现的，可能是由于低群体和/或电离过程的不可预测性。此外，尽管制备和来源不均匀，但与从单一组织和/或生物体制备的样品相比，来自商业来源的胶原样品未显示PTM的升高变化。这些发现将有助于通过MS技术获得越来越多的关于胶原蛋白PTM的信息，并最终更全面地了解胶原蛋白PTM的程度和功能作用。
Collagen is the major protein in the extracellular matrix and plays vital roles in tissue development and function. Collagen is also one of the most processed proteins in its biosynthesis. The most prominent post-translational modification (PTM) of collagen is the hydroxylation of Pro residues in the Y-position of the characteristic (Gly-Xaa-Yaa) repeating amino acid sequence of a collagen triple helix. Recent studies using mass spectrometry (MS) and tandem MS sequencing (MS/MS) have revealed unexpected hydroxylation of Pro residues in the X-positions (X-Hyp). The newly identified X-Hyp residues appear to be highly heterogeneous in location and percent occupancy. In order to understand the dynamic nature of the new X-Hyps and their potential impact on applications of MS and MS/MS for collagen research, we sampled four different collagen samples using standard MS and MS/MS techniques. We found considerable variations in the degree of PTMs of the same collagen from different organisms and/or tissues. The rat tail tendon type I collagen is particularly variable in terms of both over-hydroxylation of Pro in the X-position and under-hydroxylation of Pro in the Y-position. In contrast, only a few unexpected PTMs in collagens type I and type III from human placenta were observed. Some observations are not reproducible between different sequencing efforts of the same sample, presumably due to a low population and/or the unpredictable nature of the ionization process. Additionally, despite the heterogeneous preparation and sourcing, collagen samples from commercial sources do not show elevated variations in PTMs compared to samples prepared from a single tissue and/or organism. These findings will contribute to the growing body of information regarding the PTMs of collagen by MS technology, and culminate to a more comprehensive understanding of the extent and the functional roles of the PTMs of collagen.
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DOI：
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发表时间：
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影响因子：
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