Przybyło Małgorzata, Stepień Ewa, Pfitzner Roman, Lityńska Anna, Sadowski Jerzy
Department of Glycoconjugate Biochemistry, Institute of Zoology, Jagiellonian University, Kraków, Poland.
Arch Med Res. 2007 Jul;38(5):495-502. doi: 10.1016/j.arcmed.2007.02.001. Epub 2007 Apr 16.
The aortic valve has been the subject of many hemodynamic studies but, to our knowledge, posttranslational modification of human valve proteins has not yet been studied. Thus, the aim of this study was to determine whether any age-related changes in the protein composition of normal human aortic valves and their glycosylation pattern could be observed.
Aortic valves harvested from male cadaveric donors free of cardiovascular diseases were divided into four age groups: I, mean age 21 years; II, 30 years; III, 41 years; IV, 51 years. Proteins were separated by SDS-PAGE and transferred to PVDF membranes. Identification of monosaccharide moieties or oligosaccharide units was performed with the use of eight lectins of narrow specificity: Galantus nivalis agglutinin, Sambucus nigra agglutinin, Maackia amurensis agglutinin, Datura stramonium agglutinin, Aleuria aurantia agglutinin, Arachis hypogeae agglutinin, Phaseolus vulgaris agglutinin, and Lycopersicon esculentum agglutinin.
Isolated proteins showed no age-related changes in SDS-PAGE protein profile, contrary to their glycosylation. Protein sialylation, number of tri/tetraantennary complex glycans, proteins having terminal galactose and polylactosaminyl units increased with age, whereas protein fucosylation showed the opposite relationship. Moreover, groups III and IV possessed a larger number of proteins bearing high-mannose and/or hybrid-type glycans, and the quantity of these structures seemed to change, in particular proteins, with the age of donors.
Our results clearly demonstrate that glycosylation profile in human aortic proteins is associated with the age of the donor.
主动脉瓣一直是许多血流动力学研究的对象，但据我们所知，人类瓣膜蛋白的翻译后修饰尚未得到研究。因此，本研究的目的是确定是否能观察到正常人类主动脉瓣蛋白质组成及其糖基化模式的任何与年龄相关的变化。
从无心血管疾病的男性尸体供体获取的主动脉瓣被分为四个年龄组：I组，平均年龄21岁；II组，30岁；III组，41岁；IV组，51岁。蛋白质通过SDS-PAGE分离并转移到PVDF膜上。使用八种特异性较窄的凝集素对单糖部分或寡糖单元进行鉴定：雪花莲凝集素、黑接骨木凝集素、山嵛菜凝集素、曼陀罗凝集素、橙黄银耳凝集素、花生凝集素、菜豆凝集素和番茄凝集素。
与糖基化情况相反，分离出的蛋白质在SDS-PAGE蛋白质谱中未显示出与年龄相关的变化。蛋白质唾液酸化、三/四天线复合聚糖数量、具有末端半乳糖和多乳糖胺基单元的蛋白质随年龄增加，而蛋白质岩藻糖基化则呈现相反的关系。此外，III组和IV组含有更多带有高甘露糖和/或杂合型聚糖的蛋白质，并且这些结构的数量似乎随供体年龄变化，尤其是蛋白质。
我们的结果清楚地表明，人类主动脉蛋白中的糖基化谱与供体年龄相关。
