5ALA

Structure of Leishmania major peroxidase D211R mutant (low res)

5ALA の概要

• エントリーDOI: 10.2210/pdb5ala/pdb
• 関連するPDBエントリー: 5AL9
• 分子名称: ASCORBATE PEROXIDASE, PROTOPORPHYRIN IX CONTAINING FE, POTASSIUM ION, ... (5 entities in total)
• 機能のキーワード: oxidoreductase
• 由来する生物種: LEISHMANIA MAJOR (LEISHMANIA MAJOR)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 62624.91

構造登録者

Chreifi, G.,Hollingsworth, S.A.,Li, H.,Tripathi, S.,Arce, A.P.,Magana-Garcia, H.I.,Poulos, T.L. (登録日: 2015-03-07, 公開日: 2015-05-20, 最終更新日: 2024-01-10)

主引用文献

Chreifi, G.,Hollingsworth, S.A.,Li, H.,Tripathi, S.,Arce, A.P.,Magana-Garcia, H.I.,Poulos, T.L.
Enzymatic Mechanism of Leishmania major Peroxidase and the Critical Role of Specific Ionic Interactions.
Biochemistry, 54:3328-3336, 2015

PubMed Abstract: Leishmania major peroxidase (LmP) is very similar to the well-known yeast cytochrome c peroxidase (CcP). Both enzymes catalyze the peroxidation of cytochrome c. Like CcP, LmP reacts with H2O2 to form Compound I, which consists of a ferryl heme and a Trp radical, Fe(IV)═O;Trp(•+). Cytochrome c (Cytc) reduces the Trp radical to give Compound II, Fe(IV)═O;Trp, which is followed by an intramolecular electron transfer to give Fe(III)-OH;Trp(•+), and in the last step, Cytc reduces the Trp radical. In this study, we have used steady-state and single-turnover kinetics to improve our understanding of the overall mechanism of LmP catalysis. While the activity of CcP greatly increases with ionic strength, the kcat for LmP remains relatively constant at all ionic strengths tested. Therefore, unlike CcP, where dissociation of oxidized Cytc is limiting at low ionic strengths, association/dissociation reactions are not limiting at any ionic strength in LmP. We conclude that in LmP, the intramolecular electron transfer reaction, Fe(IV)═O;Trp to Fe(III)-OH;Trp(•+), is limiting at all ionic strengths. Unlike CcP, LmP depends on key intermolecular ion pairs to form the electron transfer competent complex. Mutating these sites causes the initial rate of association to decrease by 2 orders of magnitude and a substantial decrease in kcat. The drop in kcat is due to a switch in the rate-limiting step of the mutants from intramolecular electron transfer to the rate of association in forming the LmP-LmCytc complex. These studies show that while LmP and CcP form very similar complexes and exhibit similar activities, they substantially differ in how their activity changes as a function of ionic strength. This difference is primarily due to the heavy reliance of LmP on highly specific intermolecular ion pairs, while CcP relies mainly on nonpolar interactions.

PubMed: 25941976
DOI: 10.1021/acs.biochem.5b00338

実験手法

X-RAY DIFFRACTION (2.73 Å)