Lin, Chien-ChuChien-ChuLinSu, Shih-ChiehShih-ChiehSuSu, Ming-YuanMing-YuanSuPI-HUI LIANGFeng, Chia-ChengChia-ChengFengWu, Shih-HsiungShih-HsiungWuChang, Chung-IChung-IChang2024-03-262024-03-262016-05-0309692126https://scholars.lib.ntu.edu.tw/handle/123456789/641352The Lon AAA+ protease (LonA) is an evolutionarily conserved protease that couples the ATPase cycle into motion to drive substrate translocation and degradation. A hallmark feature shared by AAA+ proteases is the stimulation of ATPase activity by substrates. Here we report the structure of LonA bound to three ADPs, revealing the first AAA+ protease assembly where the six protomers are arranged alternately in nucleotide-free and bound states. Nucleotide binding induces large coordinated movements of conserved pore loops from two pairs of three non-adjacent protomers and shuttling of the proteolytic groove between the ATPase site and a previously unknown Arg paddle. Structural and biochemical evidence supports the roles of the substrate-bound proteolytic groove in allosteric stimulation of ATPase activity and the conserved Arg paddle in driving substrate degradation. Altogether, this work provides a molecular framework for understanding how ATP-dependent chemomechanical movements drive allosteric processes for substrate degradation in a major protein-destruction machine.enAAA+ protease; ATPase cycle; LonA; allosteric regulation; crystal structure; pore loops; protein degradation; translocation[SDGs]SDG3journal article10.1016/j.str.2016.03.001270415922-s2.0-84962091313https://api.elsevier.com/content/abstract/scopus_id/84962091313