Identification of residues involved in the interaction of Staphylococcus aureus fibronectin-binding protein with the (4)F1(5)F1 module pair of human fibronectin using heteronuclear NMR spectroscopy.
Identification of residues involved in the interaction of Staphylococcus aureus fibronectin-binding protein with the (4)F1(5)F1 module pair of human fibronectin using heteronuclear NMR spectroscopy.
复制标题
使用异核核磁共振波谱法鉴定参与金黄色葡萄球菌纤连蛋白结合蛋白与人纤连蛋白 (4)F1(5)F1 模块对相互作用的残基。
DOI：
10.1021/bi992267k
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发表时间：
2000
期刊：
影响因子：
2.9
通讯作者：
J. Potts
中科院分区：
文献类型：
--
作者：
C. Penkett;C. Dobson;L. Smith;J. R. Bright;A. Pickford;I. Campbell;J. Potts
许多致病性革兰氏阳性细菌表达与细胞外基质组分结合的细胞表面蛋白。本文研究了金黄色葡萄球菌纤连蛋白结合蛋白的配体结合重复序列（D3和D1-D4）与纤连蛋白N-末端区域的模块对（（4）F1（5）F1）之间的相互作用。当D3加入同位素标记的（4）F1（5）F1、（1）H、（15）N和（13）C NMR化学位移变化表明，结合主要是通过（4）F1中的残基，尽管（5）F1中的一些残基也受到影响。疏水和静电相互作用似乎参与。NMR数据表明，部分D3重复序列在与（4）F1（5）F1结合时从无序转变为更有序的延伸构象。在进一步的NMR实验中，在与（4）F1（5）F1结合时观察到D1-D4共振强度的选择性降低，这与先前的建议一致，即在D1、D2和D3重复序列中，主要纤连蛋白结合位点位于重复序列的C末端区域。在D1-D4中，这些区域似乎也从无序的纤连蛋白结合构象变为更有序的构象。虽然这两种蛋白质相互作用的区域以前已经被确定，但这里的发现首次确定了这两种蛋白质中可能参与相互作用的特定残基。
Many pathogenic Gram-positive bacteria express cell surface proteins that bind to components of the extracellular matrix. This paper describes studies of the interaction between ligand binding repeats (D3 and D1-D4) of a fibronectin-binding protein from Staphylococcus aureus with a module pair ((4)F1(5)F1) from the N-terminal region of fibronectin. When D3 was added to isotope-labeled (4)F1(5)F1, (1)H, (15)N, and (13)C NMR chemical shift changes indicate that binding is primarily via residues in (4)F1, although a few residues in (5)F1 are also affected. Both hydrophobic and electrostatic interactions appear to be involved. The NMR data indicate that part of the D3 repeat converts from a disordered to a more ordered, extended conformation on binding to (4)F1(5)F1. In further NMR experiments, selective reduction of the intensity of D1-D4 resonances was observed on binding to (4)F1(5)F1, consistent with previous suggestions that in each of D1, D2, and D3 repeats, the main fibronectin binding site is in the C-terminal region of the repeat. In D1-D4, these regions also appear to go from a disordered to a more ordered conformation of fibronectin binding. Although the regions of the two proteins which interact had been previously identified, the findings presented here identify, for the first time, the specific residues in both proteins that are likely to be involved in the interaction.