Skinner Andria L, Laurence Jennifer S
Department of Pharmaceutical Chemistry, The University of Kansas, Multidisciplinary Research Building Room 356, 2030 Becker Dr, Lawrence, KS 66047, USA.
Biomol NMR Assign. 2009 Jun;3(1):61-5. doi: 10.1007/s12104-008-9142-4. Epub 2009 Jan 8.
Phosphatase of regenerating liver-1 (PRL-1) is a novel target for potentially treating cancer metastases. Although its specific biochemical role in these processes has yet to be delineated, considerable evidence suggests the phosphatase activity of PRL-1 is required for promoting cancer and metastasis. PRL-1 belongs to the protein tyrosine phosphatase (PTPase) family and functions using the CX(5)R consensus active site motif. Like other PTPases, PRL-1 is inhibited by oxidation at its active site Cys, however, disulfide bond formation occurs unusually readily in wild-type PRL-1. Chemical shift assignments are available for oxidized wild type, but numerous, substantial changes are observed in the spectra upon reduction. Because the reduced form is active, we sought to identify a stable mutant that would resist oxidation and be useful for facilitating drug screening and development using NMR-based assays. We present here NMR assignments for a full-length, reduced and active form of PRL-1, PRL-1-C170S-C171S, that is well suited for this purpose.
再生肝脏磷酸酶-1（PRL-1）是潜在治疗癌症转移的新靶点。尽管其在这些过程中的具体生化作用尚未明确，但大量证据表明PRL-1的磷酸酶活性是促进癌症和转移所必需的。PRL-1属于蛋白质酪氨酸磷酸酶（PTPase）家族，利用CX(5)R共有活性位点基序发挥作用。与其他PTPases一样，PRL-1在其活性位点半胱氨酸处被氧化抑制，然而，野生型PRL-1中异常容易形成二硫键。氧化型野生型的化学位移归属是可用的，但还原后在光谱中观察到许多显著变化。由于还原形式具有活性，我们试图鉴定一种稳定的突变体，该突变体能够抵抗氧化，并有助于使用基于核磁共振的分析方法进行药物筛选和开发。我们在此展示了一种全长、还原且具有活性的PRL-1（PRL-1-C170S-C171S）的核磁共振归属，它非常适合此目的。
