Preston George W, Plusquin Michelle, Sozeri Osman, van Veldhoven Karin, Bastian Lilian, Nawrot Tim S, Chadeau-Hyam Marc, Phillips David H
MRC-PHE Centre for Environment and Health, Department of Analytical, Environmental, and Forensic Sciences, Faculty of Life Sciences and Medicine, King's College London , Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, United Kingdom.
MRC-PHE Centre for Environment and Health, Department of Epidemiology and Biostatistics, School of Public Health, Faculty of Medicine, Imperial College London , London W2 1PG, United Kingdom.
Chem Res Toxicol. 2017 Dec 18;30(12):2120-2129. doi: 10.1021/acs.chemrestox.7b00186. Epub 2017 Nov 17.
Covalently modified blood proteins (e.g., serum albumin adducts) are increasingly being viewed as potential biomarkers via which the environmental causes of human diseases may be understood. The notion that some (perhaps many) modifications have yet to be discovered has led to the development of untargeted adductomics methods, which attempt to capture entire populations of adducts. One such method is fixed-step selected reaction monitoring (FS-SRM), which analyses distributions of serum albumin adducts via shifts in the mass of a tryptic peptide [Li et al. (2011) Mol. Cell. Proteomics 10, M110.004606]. Working on the basis that FS-SRM might be able to detect biological variation due to environmental factors, we aimed to scale the methodology for use in an epidemiological setting. Development of sample preparation methods led to a batch workflow with increased throughput and provision for quality control. Challenges posed by technical and biological variation were addressed in the processing and interpretation of the data. A pilot study of 20 smokers and 20 never-smokers provided evidence of an effect of smoking on levels of putative serum albumin adducts. Differences between smokers and never-smokers were most apparent in putative adducts with net gains in mass between 105 and 114 Da (relative to unmodified albumin). The findings suggest that our implementation of FS-SRM could be useful for studying other environmental factors with relevance to human health.
共价修饰的血液蛋白质（如血清白蛋白加合物）越来越被视为潜在的生物标志物，通过它们可以了解人类疾病的环境成因。一些（或许是许多）修饰尚未被发现这一观点促使了非靶向加合物组学方法的发展，这些方法试图捕获所有类型的加合物。固定步长选择反应监测（FS-SRM）就是这样一种方法，它通过胰蛋白酶肽段质量的变化来分析血清白蛋白加合物的分布情况[Li等人（2011年），《分子与细胞蛋白质组学》10卷，M110.004606]。基于FS-SRM或许能够检测环境因素导致的生物学差异这一前提，我们旨在扩大该方法的规模以用于流行病学研究。样品制备方法的改进产生了一种批处理流程，提高了通量并具备质量控制措施。在数据处理和解读过程中解决了技术和生物学差异带来的挑战。一项针对20名吸烟者和20名从不吸烟者的初步研究提供了吸烟对假定血清白蛋白加合物水平有影响的证据。吸烟者和从不吸烟者之间的差异在质量净增加105至114道尔顿（相对于未修饰的白蛋白）的假定加合物中最为明显。研究结果表明，我们实施的FS-SRM方法可能有助于研究与人类健康相关的其他环境因素。
