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冷冻电镜和结构蛋白质组学揭示的钙调蛋白敏感腺苷酸环化酶AC8的调控位点

Regulatory sites of CaM-sensitive adenylyl cyclase AC8 revealed by cryo-EM and structural proteomics.

作者信息

Khanppnavar Basavraj, Schuster Dina, Lavriha Pia, Uliana Federico, Özel Merve, Mehta Ved, Leitner Alexander, Picotti Paola, Korkhov Volodymyr M

机构信息

Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institute, Villigen, Switzerland.

Department of Biology, Institute of Molecular Biology and Biophysics, ETH Zurich, Zurich, Switzerland.

出版信息

EMBO Rep. 2024 Mar;25(3):1513-1540. doi: 10.1038/s44319-024-00076-y. Epub 2024 Feb 13.

DOI:10.1038/s44319-024-00076-y
PMID:38351373
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10933263/
Abstract

Membrane adenylyl cyclase AC8 is regulated by G proteins and calmodulin (CaM), mediating the crosstalk between the cAMP pathway and Ca signalling. Despite the importance of AC8 in physiology, the structural basis of its regulation by G proteins and CaM is not well defined. Here, we report the 3.5 Å resolution cryo-EM structure of the bovine AC8 bound to the stimulatory Gαs protein in the presence of Ca/CaM. The structure reveals the architecture of the ordered AC8 domains bound to Gαs and the small molecule activator forskolin. The extracellular surface of AC8 features a negatively charged pocket, a potential site for unknown interactors. Despite the well-resolved forskolin density, the captured state of AC8 does not favour tight nucleotide binding. The structural proteomics approaches, limited proteolysis and crosslinking mass spectrometry (LiP-MS and XL-MS), allowed us to identify the contact sites between AC8 and its regulators, CaM, Gαs, and Gβγ, as well as to infer the conformational changes induced by these interactions. Our results provide a framework for understanding the role of flexible regions in the mechanism of AC regulation.

摘要

膜腺苷酸环化酶AC8受G蛋白和钙调蛋白（CaM）调节，介导cAMP途径与Ca信号之间的相互作用。尽管AC8在生理学中很重要，但其受G蛋白和CaM调节的结构基础尚未明确。在此，我们报道了在Ca/CaM存在下与刺激性Gαs蛋白结合的牛AC8的3.5 Å分辨率冷冻电镜结构。该结构揭示了与Gαs和小分子激活剂福斯可林结合的有序AC8结构域的架构。AC8的细胞外表面有一个带负电荷的口袋，可能是未知相互作用分子的位点。尽管福斯可林密度解析良好，但捕获的AC8状态不利于紧密核苷酸结合。结构蛋白质组学方法，即有限蛋白酶解和交联质谱（LiP-MS和XL-MS），使我们能够确定AC8与其调节因子CaM、Gαs和Gβγ之间的接触位点，并推断这些相互作用引起的构象变化。我们的数据为理解柔性区域在AC调节机制中的作用提供了一个框架。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/9a1023cf6305/44319_2024_76_Fig12_ESM.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/9a1023cf6305/44319_2024_76_Fig12_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/009f57e6d2b3/44319_2024_76_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/f0d863529310/44319_2024_76_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/b3976bbc5432/44319_2024_76_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/6bd14625ee67/44319_2024_76_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/b10993269925/44319_2024_76_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/baace22106b3/44319_2024_76_Fig8_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/6d3bf4aa789f/44319_2024_76_Fig9_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/5eb8abe02014/44319_2024_76_Fig10_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/b47d39129a37/44319_2024_76_Fig11_ESM.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/10933263/9a1023cf6305/44319_2024_76_Fig12_ESM.jpg

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2
protti: an R package for comprehensive data analysis of peptide- and protein-centric bottom-up proteomics data.Protti：一个用于以肽和蛋白质为中心的自下而上蛋白质组学数据综合数据分析的R包。
Bioinform Adv. 2021 Dec 10;2(1):vbab041. doi: 10.1093/bioadv/vbab041. eCollection 2022.
3
Structure of Cya, an evolutionary ancestor of the mammalian membrane adenylyl cyclases.
Proc Natl Acad Sci U S A. 2024 Oct 22;121(43):e2406686121. doi: 10.1073/pnas.2406686121. Epub 2024 Oct 16.
Cya 的结构，哺乳动物膜腺苷酸环化酶的进化祖先。
Elife. 2022 Aug 18;11:e77032. doi: 10.7554/eLife.77032.
4
Distinct glycerophospholipids potentiate Gsα-activated adenylyl cyclase activity.不同的甘油磷脂能增强 Gsα 激活的腺苷酸环化酶活性。
Cell Signal. 2022 Sep;97:110396. doi: 10.1016/j.cellsig.2022.110396. Epub 2022 Jul 2.
5
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6
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7
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8
Cross-Talk Between the Adenylyl Cyclase/cAMP Pathway and Ca Homeostasis.环磷酸腺苷通路与钙稳态的串扰。
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