1E30

Crystal structure of the Met148Gln mutant of rusticyanin at 1.5 Angstrom resolution

1E30 の概要

• エントリーDOI: 10.2210/pdb1e30/pdb
• 分子名称: RUSTICYANIN, COPPER (II) ION (3 entities in total)
• 機能のキーワード: rusticyanin, mutant, axial ligand, cupredoxin
• 由来する生物種: THIOBACILLUS FERROOXIDANS
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 33258.83

構造登録者

Hough, M.A.,Strange, R.W.,Hasnain, S.S. (登録日: 2000-06-02, 公開日: 2000-07-28, 最終更新日: 2025-08-13)

主引用文献

Hough, M.A.,Hall, J.F.,Kanbi, L.D.,Hasnain, S.S.
Structure of the M148Q mutant of rusticyanin at 1.5 A: a model for the copper site of stellacyanin.
Acta Crystallogr.,Sect.D, 57:355-360, 2001

PubMed Abstract: The small blue copper protein rusticyanin from Thiobacillus ferrooxidans contains a type 1 Cu centre with a single axial ligand, Met148, which together with the His-Cys-His trigonal planar ligands produces a distorted trigonal pyramidal coordination geometry to copper. Type 1 Cu sites are found in cupredoxins and several multicopper proteins, including oxidases and nitrite reductases. The role of the axial ligand has been extensively debated in terms of its function in the fine tuning of the redox potential and spectroscopic properties of type 1 Cu sites. Numerous mutations of the Met ligand in azurins have been studied, but interpretation of the results has been complicated by the presence of the additional carbonyl oxygen ligand from Gly45, a neighbouring residue to the coordinating His46. The importance of the axial ligand has been further emphasized by the finding that the type 1 centre in Rhus vernicifera stellacyanin, with the lowest redox potential in a type 1 Cu site of 184 mV, has Gln as the axial ligand, whilst fungal laccase and ceruloplasmin, which have redox potentials of 550-800 mV, have a Leu in this position. Here, the crystal structure of the M148Q mutant of rusticyanin at 1.5 A resolution is presented. This is a significantly higher resolution than that of the structures of native rusticyanin. In addition, the M148Q structure is that of the oxidized protein while the native structures to date are of the reduced protein. The mutant protein crystallizes with two molecules per asymmetric unit, in contrast to the one present in the native crystal form. This mutant's redox potential (550 mV at pH 3.2) is lowered compared with that of the native protein ( approximately 670 mV at pH 3.2) by about 120 mV. The type 1 Cu site of M148Q closely mimics the structural characteristics of the equivalent site in non-glycosylated cucumber stellacyanin (redox potential approximately 260 mV) and, owing to the absence in rusticyanin of the fifth, carbonyl ligand present in azurin, may provide a better model for the R. vernicifera stellacyanin (redox potential approximately 184 mV) type 1 Cu site, which also lacks the fifth ligand. Furthermore, the presence of two molecules in the asymmetric unit cell indicates a potential binding region of the redox partners.

PubMed: 11223511
DOI: 10.1107/s0907444900019156

実験手法

X-RAY DIFFRACTION (1.5 Å)