1YIP

Oxidized Peptidylglycine Alpha-Hydroxylating Monooxygenase (PHM) in a New Crystal Form

1YIP の概要

• エントリーDOI: 10.2210/pdb1yip/pdb
• 関連するPDBエントリー: 1OPM 1PHM 1SDW 1YI9 3PHM
• 分子名称: Peptidyl-glycine alpha-amidating monooxygenase, COPPER (II) ION (3 entities in total)
• 機能のキーワード: monooxygenase, bioactive peptide activation, copper, ascorbate, oxidoreductase
• 由来する生物種: Rattus norvegicus (Norway rat)
• 細胞内の位置: Cytoplasmic vesicle, secretory vesicle membrane; Single-pass membrane protein: P14925
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34801.85

構造登録者

Siebert, X.,Eipper, B.A.,Mains, R.E.,Prigge, S.T.,Blackburn, N.J.,Amzel, L.M. (登録日: 2005-01-12, 公開日: 2005-11-15, 最終更新日: 2024-11-13)

主引用文献

Siebert, X.,Eipper, B.A.,Mains, R.E.,Prigge, S.T.,Blackburn, N.J.,Amzel, L.M.
The Catalytic Copper of Peptidylglycine alpha-Hydroxylating Monooxygenase also Plays a Critical Structural Role.
Biophys.J., 89:3312-3319, 2005

PubMed Abstract: Many bioactive peptides require amidation of their carboxy terminus to exhibit full biological activity. Peptidylglycine alpha-hydroxylating monooxygenase (PHM; EC 1.14.17.3), the enzyme that catalyzes the first of the two steps of this reaction, is composed of two domains, each of which binds one copper atom (CuH and CuM). The CuM site includes Met(314) and two His residues as ligands. Mutation of Met(314) to Ile inactivates PHM, but has only a minimal effect on the EXAFS spectrum of the oxidized enzyme, implying that it contributes only marginally to stabilization of the CuM site. To characterize the role of Met(314) as a CuM ligand, we determined the structure of the Met(314)Ile-PHM mutant. Since the mutant protein failed to crystallize in the conditions of the original wild-type protein, this structure determination required finding a new crystal form. The Met(314)Ile-PHM mutant structure confirms that the mutation does not abolish CuM binding to the enzyme, but causes other structural perturbations that affect the overall stability of the enzyme and the integrity of the CuH site. To eliminate possible effects of crystal contacts, we redetermined the structure of wt-PHM in the Met(314)Ile-PHM crystal form and showed that it does not differ from the structure of wild-type (wt)-PHM in the original crystals. Met(314)Ile-PHM was also shown to be less stable than wt-PHM by differential scanning calorimetry. Both structural and calorimetric studies point to a structural role for the CuM site, in addition to its established catalytic role.

PubMed: 16100265
DOI: 10.1529/biophysj.105.066100

実験手法

X-RAY DIFFRACTION (2.2 Å)