Zhou M M, Logan T M, Thèriault Y, Van Etten R L, Fesik S W
Pharmaceutical Discovery Division, Abbott Laboratories, Abbott Park, Illinois 60064.
Biochemistry. 1994 May 3;33(17):5221-9. doi: 10.1021/bi00183a027.
Phosphotyrosyl protein phosphatases play an important role in mediating cellular signal transduction; yet three-dimensional structures of this important class of proteins have not been reported. We present the sequence-specific 1H, 13C, and 15N backbone assignments for the low molecular weight bovine heart phosphotyrosyl protein phosphatase (BHPTPase) (157 residues, 17,900). The assignments were obtained from a combination of double- and triple-resonance multidimensional NMR experiments. From these assignments, the secondary structure of BHPTPase was determined from an analysis of NOE patterns, 3JHNH alpha coupling constants, 13C alpha and 13CO chemical shifts, and amide 1H exchange rates. BHPTPase was found to consist of a four-stranded parallel beta-sheet (residues K6-C12, W39-A45, Y87-M91, and K112-L116), four alpha-helices (residues I21-D32, R58-G67, S94-N104, and D135-R157), and one stretch of beta 10-helix (residues K79-F85). The secondary structure is characteristic of the beta alpha beta structural motif. The secondary structure elements identified in this study are consistent with previous chemical and mutagenesis studies of BHPTPase structure.
磷酸酪氨酸蛋白磷酸酶在介导细胞信号转导中发挥着重要作用；然而，这类重要蛋白质的三维结构尚未见报道。我们给出了低分子量牛心磷酸酪氨酸蛋白磷酸酶（BHPTPase）（157个残基，17,900）的序列特异性1H、13C和15N主链归属。这些归属是通过双共振和三共振多维核磁共振实验相结合获得的。基于这些归属，通过对核Overhauser效应（NOE）模式、3JHNHα耦合常数、13Cα和13CO化学位移以及酰胺1H交换率的分析，确定了BHPTPase的二级结构。结果发现，BHPTPase由一个四链平行β折叠（残基K6 - C12、W39 - A45、Y87 - M91和K112 - L116）、四个α螺旋（残基I21 - D32、R58 - G67、S94 - N104和D135 - R157）以及一段β10螺旋（残基K79 - F85）组成。其二级结构具有βαβ结构基序的特征。本研究中鉴定出的二级结构元件与先前关于BHPTPase结构的化学和诱变研究结果一致。
