Peterson Janine H, Woolhead Cheryl A, Bernstein Harris D
Genetics and Biochemistry Branch, NIDDK, National Institutes of Health, Bethesda, Maryland 20892-0538, USA.
J Biol Chem. 2003 Nov 14;278(46):46155-62. doi: 10.1074/jbc.M309082200. Epub 2003 Aug 29.
Previous studies have demonstrated that signal peptides bind to the signal recognition particle (SRP) primarily via hydrophobic interactions with the 54-kDa protein subunit. The crystal structure of the conserved SRP ribonucleoprotein core, however, raised the surprising possibility that electrostatic interactions between basic amino acids in signal peptides and the phosphate backbone of SRP RNA may also play a role in signal sequence recognition. To test this possibility we examined the degree to which basic amino acids in a signal peptide influence the targeting of two Escherichia coli proteins, maltose binding protein and OmpA. Whereas both proteins are normally targeted to the inner membrane by SecB, we found that replacement of their native signal peptides with another moderately hydrophobic but unusually basic signal peptide (DeltaEspP) rerouted them into the SRP pathway. Reduction in either the net positive charge or the hydrophobicity of the DeltaEspP signal peptide decreased the effectiveness of SRP recognition. A high degree of hydrophobicity, however, compensated for the loss of basic residues and restored SRP binding. Taken together, the data suggest that the formation of salt bridges between SRP RNA and basic amino acids facilitates the binding of a distinct subset of signal peptides whose hydrophobicity falls slightly below a threshold level.
先前的研究表明,信号肽主要通过与54 kDa蛋白质亚基的疏水相互作用与信号识别颗粒(SRP)结合。然而,保守的SRP核糖核蛋白核心的晶体结构提出了一种惊人的可能性,即信号肽中的碱性氨基酸与SRP RNA的磷酸骨架之间的静电相互作用也可能在信号序列识别中发挥作用。为了验证这种可能性,我们研究了信号肽中的碱性氨基酸对两种大肠杆菌蛋白——麦芽糖结合蛋白和OmpA靶向作用的影响程度。虽然这两种蛋白通常通过SecB靶向内膜,但我们发现用另一种适度疏水但异常碱性的信号肽(DeltaEspP)替换它们的天然信号肽会将它们重新导向SRP途径。DeltaEspP信号肽的净正电荷或疏水性的降低会降低SRP识别的有效性。然而,高度的疏水性补偿了碱性残基的损失并恢复了SRP结合。综上所述,数据表明SRP RNA与碱性氨基酸之间形成盐桥有助于疏水性略低于阈值水平的特定信号肽子集的结合。