title: Chaperonin GroEL-GroES Functions as both Alternating and Non-Alternating Engines.
abstract: A double ring-shaped GroEL consisting of 14 ATPase subunits assists protein folding, together with co-chaperonin GroES. The dynamic GroEL-GroES interaction is actively involved in the chaperonin reaction. Therefore, revealing this dynamic interaction is a key to understanding the operation principle of GroEL. Nevertheless, how this interaction proceeds in the reaction cycle has long been controversial. Here, we directly imaged GroEL-GroES interaction in the presence of disulfide-reduced α-lactalbumin as a substrate protein using high-speed atomic force microscopy. This real-time imaging revealed the occurrence of primary, symmetric GroEL:GroES2 and secondary, asymmetric GroEL:GroES1 complexes. Remarkably, the reaction was observed to often branch into main and side pathways. In the main pathway, alternate binding and release of GroES occurs at the two rings, indicating tight cooperation between the two rings. In the side pathway, however, this cooperation is disrupted, resulting in the interruption of alternating rhythm. From various properties observed for both pathways, we provide mechanistic insight into the alternate and non-alternate operations of the two-engine system.
authors.affiliation: Department of Physics, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan; CREST, JST, Sanban-cho, Chiyoda-ku, Tokyo 102-0075, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Yamamoto
authors.affiliation: Department of Physics, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan; CREST, JST, Sanban-cho, Chiyoda-ku, Tokyo 102-0075, Japan. Electronic address: tando@staff.kanazawa-u.ac.jp.
authors.forename: Toshio
authors.initials: T
authors.lastname: Ando
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D018835:Chaperonin 10 / Q000378:metabolism*; D018834:Chaperonin 60 / Q000378:metabolism*; D018833:Chaperonins / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D007768:Lactalbumin / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D017510:Protein Folding
chemical_list: D018835:Chaperonin 10; D018834:Chaperonin 60; D007768:Lactalbumin; D000251:Adenosine Triphosphatases; D018833:Chaperonins
keywords: GroEL–GroES interaction; chaperonin; cooperativity; high-speed AFM