5CAA

Structure of Leishmania nucleoside diphosphate kinase mutant P100S/del5-Cterm

5CAA の概要

• エントリーDOI: 10.2210/pdb5caa/pdb
• 関連するPDBエントリー: 3NGS 5C7P 5CAB
• 分子名称: Nucleoside diphosphate kinase (2 entities in total)
• 機能のキーワード: leishmania major, nucleoside diphosphate kinase, site directed-mutagenesis, quaternary structure, conformational stability, transferase
• 由来する生物種: Leishmania major
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 36403.26

構造登録者

Vieira, P.S.,de Giuseppe, P.O.,de Oliveira, A.H.C.,Murakami, M.T. (登録日: 2015-06-29, 公開日: 2015-10-14, 最終更新日: 2023-09-27)

主引用文献

Vieira, P.S.,de Giuseppe, P.O.,de Oliveira, A.H.,Murakami, M.T.
The role of the C-terminus and Kpn loop in the quaternary structure stability of nucleoside diphosphate kinase from Leishmania parasites.
J.Struct.Biol., 192:336-341, 2015

PubMed Abstract: Nucleoside diphosphate kinase (NDK) is a housekeeping enzyme that plays key roles in nucleotide recycling and homeostasis in trypanosomatids. Moreover, it is secreted by the intracellular parasite Leishmania to modulate the host response. These functions make NDK an attractive target for drug design and for studies aiming at a better understanding of the mechanisms mediating host-pathogen interactions. Here, we report the crystal structures of three mutants of the NDK from Leishmania major (LmNDK) that affects the stability of the hexameric biological assembly including P95S, Δ5Ct (lacking the last five residues) and the double mutant P100S/Δ5Ct. Although P95S and Δ5Ct variants conserve the hexameric structure of the wild-type protein, the double mutant becomes a dimer as shown by in solution studies. Free energy calculation of dimer-dimer interfaces and enzymatic assays indicate that P95S, Δ5Ct and P100S/Δ5Ct mutations progressively decrease the hexamer stability and enzyme activity. These results demonstrate that the mutated regions play a role in protein function through stabilizing the quaternary arrangement.

PubMed: 26410384
DOI: 10.1016/j.jsb.2015.09.009

実験手法

X-RAY DIFFRACTION (2.3 Å)