Department of Biochemistry, Memorial University of Newfoundland, St. John's, Newfoundland and Labrador, Canada.
PLoS One. 2013 Sep 2;8(9):e72821. doi: 10.1371/journal.pone.0072821. eCollection 2013.
Breathing is enabled by lung surfactant, a mixture of proteins and lipids that forms a surface-active layer and reduces surface tension at the air-water interface in lungs. Surfactant protein B (SP-B) is an essential component of lung surfactant. In this study we probe the mechanism underlying the important functional contributions made by the N-terminal 7 residues of SP-B, a region sometimes called the "insertion sequence". These studies employed a construct of SP-B, SP-B (1-25,63-78), also called Super Mini-B, which is a 41-residue peptide with internal disulfide bonds comprising the N-terminal 7-residue insertion sequence and the N- and C-terminal helices of SP-B. Circular dichroism, solution NMR, and solid state (2)H NMR were used to study the structure of SP-B (1-25,63-78) and its interactions with phospholipid bilayers. Comparison of results for SP-B (8-25,63-78) and SP-B (1-25,63-78) demonstrates that the presence of the 7-residue insertion sequence induces substantial disorder near the centre of the lipid bilayer, but without a major disruption of the overall mechanical orientation of the bilayers. This observation suggests the insertion sequence is unlikely to penetrate deeply into the bilayer. The 7-residue insertion sequence substantially increases the solution NMR linewidths, most likely due to an increase in global dynamics.
呼吸是由肺表面活性剂(一种蛋白质和脂质的混合物)实现的,它形成一个表面活性层,降低了肺部气-水界面的表面张力。表面活性蛋白 B(SP-B)是肺表面活性剂的重要组成部分。在这项研究中,我们探讨了 SP-B 的 N 端 7 个残基(有时称为“插入序列”)发挥重要功能的机制。这些研究采用了 SP-B 的一种构建体,即 SP-B(1-25,63-78),也称为超级 Mini-B,它是一个 41 个残基的肽,具有内部二硫键,由 N 端 7 个残基的插入序列和 SP-B 的 N 和 C 端螺旋组成。圆二色性、溶液 NMR 和固态(2)H NMR 用于研究 SP-B(1-25,63-78)的结构及其与磷脂双层的相互作用。比较 SP-B(8-25,63-78)和 SP-B(1-25,63-78)的结果表明,插入序列的存在在脂质双层的中心附近诱导了大量的无序,但没有对双层的整体机械取向造成重大破坏。这一观察结果表明,插入序列不太可能深入双层。7 个残基的插入序列大大增加了溶液 NMR 的线宽,这很可能是由于整体动力学的增加。