Hogarth Caroline J, Fitzpatrick Julie L, Nolan Andrea M, Young Fiona J, Pitt Andrew, Eckersall P David
Department of Veterinary Clinical Studies, Centre for Integrated Diagnostic Systems, University of Glasgow, Thomson Building, Gilmorehill, Glasgow G12 8QQ, Scotland, UK.
Proteomics. 2004 Jul;4(7):2094-100. doi: 10.1002/pmic.200300723.
During clinical mastitis in dairy cows, the quantity of milk produced decreases and the composition of the milk is altered. As the severity of inflammation associated with the disease increases, the chemical composition of milk approaches that of blood as a consequence of increased permeability of the blood mammary barrier, or de novo intramammary synthesis, as has been suggested for mammary associated serum amyloid A3. A better understanding of these events may provide new approaches for the diagnosis and treatment of mastitis. The objective of this study was to document the changes in the protein composition of milk during clinical mastitis using a proteomic approach, with the objective of identifying new diagnostic markers of mastitis. Whey from dairy cows with clinical mastitis was compared to whey from healthy animals by two-dimensional gel electrophoresis (2-DE) with colloidal Coomassie staining and matrix-assisted desorption/ionization mass spectrometry. Increases in the concentrations of proteins of blood serum origin, including serotransferrin and albumin, were identified in mastitic whey compared to normal whey, while concentrations of the major whey proteins alpha-lactalbumin and beta-lactoglobulin were reduced in mastitic whey. Mass spectrometry subsequently confirmed the location of albumin, alpha-lactalbumin and beta-lactoglobulin on the 2-DE gels at M(r)/pI of 69 294/5.8, 14 200/4.5 and 19 883/4.9 respectively.
在奶牛临床型乳腺炎期间,产奶量会下降,且牛奶成分会发生改变。随着与该疾病相关的炎症严重程度增加,由于血乳屏障通透性增加或乳腺内从头合成(如乳腺相关血清淀粉样蛋白A3所提示的那样),牛奶的化学成分会接近血液。更好地理解这些事件可能为乳腺炎的诊断和治疗提供新方法。本研究的目的是使用蛋白质组学方法记录临床型乳腺炎期间牛奶蛋白质组成的变化,以识别乳腺炎的新诊断标志物。通过二维凝胶电泳(2-DE)结合胶体考马斯亮蓝染色和基质辅助激光解吸/电离质谱,将患有临床型乳腺炎的奶牛的乳清与健康动物的乳清进行比较。与正常乳清相比,在患乳腺炎的乳清中发现血清来源蛋白质(包括转铁蛋白和白蛋白)的浓度增加,而主要乳清蛋白α-乳白蛋白和β-乳球蛋白的浓度在患乳腺炎的乳清中降低。质谱随后证实了白蛋白、α-乳白蛋白和β-乳球蛋白在2-DE凝胶上的位置,其相对分子质量/等电点分别为69294/5.8、14200/4.5和19883/4.9。