Bhat Vadiraja B, Choi Man Ho, Wishnok John S, Tannenbaum Steven R
Biological Engineering Division and Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, 56-731A, Cambridge, MA 02139, USA.
J Proteome Res. 2005 Sep-Oct;4(5):1814-25. doi: 10.1021/pr0501463.
In SJL mice, growth of RcsX lymphoma cells induces an inflammatory response by stimulating V(beta)16+ T cells. During inflammation, various serum protein levels can increase (e.g., acute phase reactants) or decrease (e.g., albumin), and most of these altered proteins are thus potential biomarkers. Although blood plasma is a valuable and promising sample for biomarker discovery for diseases or for novel drug targets, its proteome is complex. To address this, we have focused on a comprehensive comparison of the plasma proteomes from normal and RcsX-tumor-bearing SJL mice using the 1D-Gel-LC-MS/MS method after removing albumin and immunoglobulins. This analysis resulted in the identification of a total of 1079 nonredundant mouse plasma proteins; more than 480 in normal and 790 in RcsX-tumor-bearing SJL mouse plasma. Of these, only 191 proteins were found in common. The molecular weights ranged from 2 to 876 kDa, covering the pI values between 4.22 and 12.09, and included proteins with predicted transmembrane domains. By comparing the plasma proteomic profile of normal and RcsX-tumor-bearing SJL mice, we found significant changes in the levels of many proteins in RcsX-tumor-bearing mouse plasma. Most of the up-regulated proteins were identified as acute-phase proteins (APPs). Also, several unique proteins i.e., haptoglobin, proteosome subunits, fetuin-B, 14-3-3 zeta, MAGE-B4 antigen, etc, were found only in the tumor-bearing mouse plasma; either secreted, shed by membrane vesicles, or externalized due to cell death. These results affirm the effectiveness of this approach for protein identification from small samples, and for comparative proteomics in potential animal models of human disorders.
在SJL小鼠中，RcsX淋巴瘤细胞的生长通过刺激V（β）16 + T细胞诱导炎症反应。在炎症过程中，各种血清蛋白水平可能会升高（如急性期反应物）或降低（如白蛋白），因此这些改变的蛋白大多是潜在的生物标志物。尽管血浆是用于发现疾病生物标志物或新型药物靶点的有价值且有前景的样本，但其蛋白质组很复杂。为了解决这个问题，我们专注于在去除白蛋白和免疫球蛋白后，使用一维凝胶液相色谱 - 串联质谱法对正常SJL小鼠和携带RcsX肿瘤的SJL小鼠的血浆蛋白质组进行全面比较。该分析共鉴定出1079种非冗余小鼠血浆蛋白；正常SJL小鼠血浆中有480多种，携带RcsX肿瘤的SJL小鼠血浆中有790多种。其中，仅发现191种蛋白是共有的。分子量范围为2至876 kDa，涵盖4.22至12.09的pI值，并且包括具有预测跨膜结构域的蛋白。通过比较正常SJL小鼠和携带RcsX肿瘤的SJL小鼠的血浆蛋白质组图谱，我们发现在携带RcsX肿瘤的小鼠血浆中许多蛋白的水平有显著变化。大多数上调的蛋白被鉴定为急性期蛋白（APPs）。此外，还发现了几种独特的蛋白，即触珠蛋白、蛋白酶体亚基、胎球蛋白 - B、14 - 3 - 3 ζ、MAGE - B4抗原等，仅在携带肿瘤的小鼠血浆中存在；它们要么是分泌的，通过膜泡脱落的，要么是由于细胞死亡而外化的。这些结果证实了这种方法对于从小样本中鉴定蛋白质以及在人类疾病潜在动物模型中进行比较蛋白质组学的有效性。
