Structure of Metal Site in Azurin, Met121 Mutants of Azurin, and Stellacyanin Investigated by 111mCd Perturbed Angular Correlation (PAC) (∗)

The geometries of the metal sites in cadmium-substituted azurins have been investigated by 111mCd perturbed angular correlation (PAC). The study includes wild type azurin as well as Met121 mutants of azurin, where methionine has been substituted by Ala, Asn, Asp, Gln, Glu, and Leu. The nuclear quadr...

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Bibliographic Details
Published in:The Journal of biological chemistry 1995-01, Vol.270 (2), p.573-580
Main Authors: Danielsen, Eva, Bauer, Rogert, Hemmingsen, Lars, Andersen, Marie-Louise, Bjerrum, Morten J., Butz, Tilman, Tröger, Wolfgang, Canters, Gerard W., Hoitink, Carla W.G., Karlsson, Göran, Hansson, Örjan, Messerschmidt, Albrecht
Format: Article
Language:English
Subjects:
Azurin - chemistry
Azurin - genetics
Azurin - metabolism
Cadmium - metabolism
Fourier Analysis
Indium Radioisotopes
Metalloproteins - chemistry
Metalloproteins - metabolism
Methionine - metabolism
Mutation
Plant Proteins - chemistry
Plant Proteins - metabolism
Spectrum Analysis
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Summary:The geometries of the metal sites in cadmium-substituted azurins have been investigated by 111mCd perturbed angular correlation (PAC). The study includes wild type azurin as well as Met121 mutants of azurin, where methionine has been substituted by Ala, Asn, Asp, Gln, Glu, and Leu. The nuclear quadrupole interaction of wild type azurin analyzed in the angular overlap model is well described as coordination of His46, His117, and Cys112 and cannot be described by coordination of Met121 and/or Gly45. For most of the mutants, there exist two coordination geometries of the cadmium ion. With the exception of the Glu and Asp mutants, one of the conformations is similar to the wild type conformation. The other coordination geometries are either best described by a coordinating water molecule close to the original methionine position or by coordination by the substituting amino acid. These experiments show that even though the methionine does not coordinate it plays an important role for the geometry of the metal site. The nuclear quadrupole interaction of stellacyanin was also measured. The value resembles the most prominent nuclear quadrupole interaction of the Met121121→ Gln mutant of Alcaligenes denitrificans azurin, indicating that the structures of the two metal sites are similar.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.270.2.573