1Q99

Crystal structure of the Saccharomyces cerevisiae SR protein kinsae, Sky1p, complexed with the non-hydrolyzable ATP analogue, AMP-PNP

1Q99 の概要

• エントリーDOI: 10.2210/pdb1q99/pdb
• 関連するPDBエントリー: 1q8y 1q8z 1q97
• 分子名称: SR protein kinsae, NICKEL (II) ION, SULFATE ION, ... (7 entities in total)
• 機能のキーワード: protein kinase, transferase
• 由来する生物種: Saccharomyces cerevisiae (baker's yeast); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 87560.26

構造登録者

Nolen, B.,Ngo, J.,Chakrabarti, S.,Vu, D.,Adams, J.A.,Ghosh, G. (登録日: 2003-08-22, 公開日: 2003-09-23, 最終更新日: 2024-02-14)

主引用文献

Nolen, B.,Ngo, J.,Chakrabarti, S.,Vu, D.,Adams, J.A.,Ghosh, G.
Nucleotide-Induced Conformational Changes in the Saccharomyces cerevisiae SR Protein Kinase, Sky1p, Revealed by X-ray Crystallography
Biochemistry, 42:9575-9585, 2003

PubMed Abstract: Conformational changes are thought to play a key role in the function of active protein kinases, although little is known about how these changes relate to the mechanism of phosphorylation. Here we present four high-resolution structures of a single crystal form of Sky1p, a constitutively active serine kinase implicated in yeast RNA processing, each in a different state of nucleotide binding. By comparing the apoenzyme structure to the ADP- and ATP-bound Sky1p structures, we have revealed conformational changes caused by ATP binding or conversion from nucleotide reactant to product. Rotation of the small lobe of the kinase closes the cleft upon binding, allowing the nucleotide to interact with residues from both lobes of the kinase, although some interactions thought to be important for phosphotransfer are missing in the ATP-containing structure. In the apoenzyme, a kinase-conserved phosphate-anchoring loop is in a twisted conformation that is incompatible with ADP and ATP binding, providing a potential mechanism for facilitating ADP release in Sky1p. The nonhydrolyzable ATP analogue AMP-PNP binds in a unique mode that fails to induce lobe closure. This observation, along with comparisons between the two independent molecules in the asymmetric unit of each structure, has provided new molecular details about how the nucleotide binds and induces closure. Finally, we have used mutational analysis to establish the importance of a glycine within the linker that connects the two lobes of Sky1p.

PubMed: 12911299
DOI: 10.1021/bi0344331

実験手法

X-RAY DIFFRACTION (2.11 Å)