Institute of Pharmaceutical Sciences, Pharmaceutical Chemistry, University of Graz, 8010 Graz, Austria.
BioTechMed-Graz, 8010 Graz, Austria.
J Proteome Res. 2023 Jun 2;22(6):1790-1799. doi: 10.1021/acs.jproteome.2c00795. Epub 2023 Apr 13.
BCR-ABL is the oncogenic fusion product of tyrosine kinase ABL1 and a highly frequent driver of acute lymphocytic leukemia (ALL) and chronic myeloid leukemia (CML). The kinase activity of BCR-ABL is strongly elevated; however, changes of substrate specificity in comparison to wild-type ABL1 kinase are less well characterized. Here, we heterologously expressed full-length BCR-ABL kinases in yeast. We exploited the proteome of living yeast as an phospho-tyrosine substrate for assaying human kinase specificity. Phospho-proteomic analysis of ABL1 and BCR-ABL isoforms p190 and p210 yielded a high-confidence data set of 1127 phospho-tyrosine sites on 821 yeast proteins. We used this data set to generate linear phosphorylation site motifs for ABL1 and the oncogenic ABL1 fusion proteins. The oncogenic kinases yielded a substantially different linear motif when compared to ABL1. Kinase set enrichment analysis with human pY-sites that have high linear motif scores well-recalled BCR-ABL driven cancer cell lines from human phospho-proteome data sets.
BCR-ABL 是酪氨酸激酶 ABL1 的致癌融合产物，是急性淋巴细胞白血病（ALL）和慢性髓性白血病（CML）的高频驱动因素。BCR-ABL 的激酶活性显著升高；然而，与野生型 ABL1 激酶相比，其底物特异性的变化特征描述较少。在这里，我们在酵母中异源表达全长 BCR-ABL 激酶。我们利用活酵母的蛋白质组作为检测人激酶特异性的磷酸酪氨酸底物。ABL1 和 BCR-ABL 同工型 p190 和 p210 的磷酸蛋白质组分析产生了一个由 821 种酵母蛋白上的 1127 个磷酸酪氨酸位点组成的高可信度数据集。我们使用该数据集为 ABL1 和致癌 ABL1 融合蛋白生成线性磷酸化位点基序。与 ABL1 相比，致癌激酶产生了一个明显不同的线性基序。当对具有高线性基序评分的人类 pY 位点进行激酶集富集分析时，这些位点很好地召回了来自人类磷酸蛋白质组数据集中的 BCR-ABL 驱动的癌细胞系。
