通过中子衍射确定细菌视紫红质结构中化学修饰赖氨酸 41 的位置。

Location of chemically modified lysine 41 in the structure of bacteriorhodopsin by neutron diffraction.

出版信息

Biophys J. 1986 Oct;50(4):629-35. doi: 10.1016/S0006-3495(86)83502-0.

DOI:10.1016/S0006-3495(86)83502-0

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1329840/

Abstract

Purple membranes were prepared in which bacteriorhodopsin was labeled at lysine 41 with phenylisothiocyanate (PITC) and with perdeuterated PITC. The in-plane position of this small label containing only five deuterons was determined from the differences between the neutron diffraction intensities of the two samples. At 8.7-A resolution the Fourier difference map revealed a well-defined site between helices 3 and 4. This position was confirmed by a refinement procedure in reciprocal space. Model calculations showed that the observed difference density had the right amplitude for the label. Thus it is possible to locate a small group in a large protein structure by replacing as few as five hydrogens by deuterium. The observed location of PITC restricts the number of possibilities for the assignment of helix B in the sequence (to which lysine 41 is attached) to one of the seven helices of the structure. Taking into account the size of the label and the length of the lysine side chain our result excludes helices 1, 2, and 7 as candidates for B.

摘要

紫色膜中，细菌视紫红质的赖氨酸 41 被苯异硫氰酸酯（PITC）和氘代 PITC 标记。通过比较这两个样品的中子衍射强度差异，确定了仅含有五个氘原子的这个小标记的面内位置。在 8.7-A 的分辨率下，傅里叶差图揭示了在 3 号和 4 号螺旋之间有一个明确的位置。该位置通过倒空间的精修过程得到了证实。模型计算表明，观察到的差异密度对标记具有正确的幅度。因此，通过用氘取代多达五个氢原子，就有可能在大型蛋白质结构中定位一个小基团。PITC 的观察位置限制了序列中赖氨酸 41 所连接的螺旋 B 的分配的可能性，只有结构的七个螺旋之一。考虑到标记的大小和赖氨酸侧链的长度，我们的结果排除了 1、2 和 7 号螺旋作为 B 的候选者。

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