通过交联/质谱法获得牛视网膜棒状外节制剂全长鸟苷酸环化酶 1 的首个 3D 结构数据。

First 3D-Structural Data of Full-Length Guanylyl Cyclase 1 in Rod-Outer-Segment Preparations of Bovine Retina by Cross-Linking/Mass Spectrometry.

机构信息

Department of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, Halle/Saale, Germany.

Interdisciplinary Research Center HALOmem, Charles Tanford Protein Center, Martin Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3a, Halle/Saale, Germany; Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, Halle/Saale, Germany; Biozentrum, Martin Luther University Halle-Wittenberg, Weinbergweg 22, Halle/Saale, Germany.

出版信息

J Mol Biol. 2021 May 14;433(10):166947. doi: 10.1016/j.jmb.2021.166947. Epub 2021 Mar 17.

DOI:10.1016/j.jmb.2021.166947

PMID:33744315

Abstract

The rod-outer-segment guanylyl cyclase 1 (ROS-GC1) is a key transmembrane protein for retinal phototransduction. Mutations of ROS-GC1 correlate with different retinal diseases that often lead to blindness. No structural data are available for ROS-GC1 so far. We performed a 3D-structural analysis of native ROS-GC1 from bovine retina by cross-linking/mass spectrometry (XL-MS) and computational modeling. Absolute quantification and activity measurements of native ROS-GC1 were performed by MS-based assays directly in bovine retina samples. Our data present the first 3D-structural analysis of active, full-length ROS-GC1 derived from bovine retina. We propose a novel domain organization for the intracellular domain ROS-GC1. Our XL-MS data of native ROS-GC1 from rod-outer-segment preparations of bovine retina agree with a dimeric architecture. Our integrated approach can serve as a blueprint for conducting 3D-structural studies of membrane proteins in their native environment.

摘要

棒体细胞外环鸟苷酸环化酶 1（ROS-GC1）是视网膜光转导的关键跨膜蛋白。ROS-GC1 的突变与多种常导致失明的视网膜疾病相关。目前尚无 ROS-GC1 的结构数据。我们通过交联/质谱（XL-MS）和计算建模对牛视网膜中的天然 ROS-GC1 进行了 3D 结构分析。通过 MS 为基础的检测方法直接在牛视网膜样本中进行天然 ROS-GC1 的绝对定量和活性测量。我们的数据提供了来自牛视网膜的活性全长 ROS-GC1 的首个 3D 结构分析。我们提出了一种新的 ROS-GC1 胞内域的结构组织。我们对来自牛视网膜棒体细胞外节的天然 ROS-GC1 的 XL-MS 数据与二聚体结构一致。我们的综合方法可以作为在天然环境中对膜蛋白进行 3D 结构研究的蓝图。