5OOW

Crystal structure of lobe II from the nucleotide binding domain of DnaK in complex with AMPPCP

5OOW の概要

• エントリーDOI: 10.2210/pdb5oow/pdb
• 分子名称: Chaperone protein DnaK, PHOSPHOMETHYLPHOSPHONIC ACID ADENYLATE ESTER (3 entities in total)
• 機能のキーワード: hsp70, nucleotide binding domain, nbd, sugar kinase family, chaperone
• 由来する生物種: Escherichia coli (strain K12)
• 細胞内の位置: Cytoplasm : P0A6Y8
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 45927.41

構造登録者

Jakob, R.P.,Bauer, D.,Meinhold, S.,Stigler, J.,Merkel, U.,Maier, T.,Rief, M.,Zoldak, G. (登録日: 2017-08-09, 公開日: 2018-04-25, 最終更新日: 2024-01-17)

主引用文献

Bauer, D.,Meinhold, S.,Jakob, R.P.,Stigler, J.,Merkel, U.,Maier, T.,Rief, M.,Zoldak, G.
A folding nucleus and minimal ATP binding domain of Hsp70 identified by single-molecule force spectroscopy.
Proc. Natl. Acad. Sci. U.S.A., 115:4666-4671, 2018

PubMed Abstract: The folding pathways of large proteins are complex, with many of them requiring the aid of chaperones and others folding spontaneously. Along the folding pathways, partially folded intermediates are frequently populated; their role in the driving of the folding process is unclear. The structures of these intermediates are generally not amenable to high-resolution structural techniques because of their transient nature. Here we employed single-molecule force measurements to scrutinize the hierarchy of intermediate structures along the folding pathway of the nucleotide binding domain (NBD) of Hsp70 DnaK. DnaK-NBD is a member of the sugar kinase superfamily that includes Hsp70s and the cytoskeletal protein actin. Using optical tweezers, a stable nucleotide-binding competent folding intermediate comprising lobe II residues (183-383) was identified as a critical checkpoint for productive folding. We obtained a structural snapshot of this folding intermediate that shows native-like conformation. To assess the fundamental role of folded lobe II for efficient folding, we turned our attention to yeast mitochondrial NBD, which does not fold without a dedicated chaperone. After replacing the yeast lobe II residues with stable lobe II, the obtained chimeric protein showed native-like ATPase activity and robust folding into the native state, even in the absence of chaperone. In summary, lobe II is a stable nucleotide-binding competent folding nucleus that is the key to time-efficient folding and possibly resembles a common ancestor domain. Our findings provide a conceptual framework for the folding pathways of other members of this protein superfamily.

PubMed: 29669923
DOI: 10.1073/pnas.1716899115

実験手法

X-RAY DIFFRACTION (2.9 Å)