优化骨骼肌组织和细胞中线粒体、内质网或线粒体-内质网接触位点（MERC）标志物的原位邻近连接分析

Optimizing In Situ Proximity Ligation Assays for Mitochondria, ER, or MERC Markers in Skeletal Muscle Tissue and Cells.

作者信息

Crabtree Amber, Le Han, Harris Chanel, Oliver Ashton, Barillas Andy, Moore Johnathan, Nguyen Desiree Ngoc-Ha, Rabago Izabella Marie, Rodriguez Benjamin, Stephens Dominique C, Beasley Heather K, Garza-Lopez Edgar, Neikirk Kit, Mungai Margaret, Vang Larry, Vue Zer, Vue Neng, Marshall Andrea G, Turner Kyrin, Shao Jianqiang, Murray Sandra, Gaddy Jennifer A, Wanjalla Celestine, Davis Jamaine, Damo Steven, Hinton Antentor O

机构信息

Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee.

These authors contributed equally to this work.

出版信息

Curr Protoc. 2025 Feb;5(2):e70043. doi: 10.1002/cpz1.70043.

DOI:10.1002/cpz1.70043

PMID:39906960

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11795341/

Abstract

Proximity ligation assays (PLAs) use specific antibodies to detect endogenous protein-protein interactions. PLAs are a highly useful biochemical technique that allow two proteins within proximity to be visualized with fluorescent probes amplified by PCR. While this technique has gained prominence, the use of a PLA in mouse skeletal muscle (SkM) is novel. In this article, we discuss how the PLA method can be used in SkM to study the protein-protein interactions within mitochondria-endoplasmic reticulum contact sites (MERCs). © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Proximity ligation assay for skeletal muscle tissue and myoblast for MERC proteins.

摘要

邻近连接分析（PLA）使用特异性抗体来检测内源性蛋白质-蛋白质相互作用。PLA是一种非常有用的生化技术，它能使邻近的两种蛋白质通过PCR扩增的荧光探针进行可视化。虽然这项技术已备受关注，但在小鼠骨骼肌（SkM）中使用PLA还是新颖的。在本文中，我们讨论了如何将PLA方法用于SkM，以研究线粒体-内质网接触位点（MERC）内的蛋白质-蛋白质相互作用。© 2025作者。由Wiley Periodicals LLC出版的《当前方案》。基本方案：用于骨骼肌组织和成肌细胞中MERC蛋白的邻近连接分析。