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Sulfolobus acidocaldarius inorganic pyrophosphatase: structure, thermostability, and effect of metal ion in an archael pyrophosphatase.
Leppänen, V M; Nummelin, H; Hansen, T; Lahti, R; Schäfer, G; Goldman, A.
Afiliación
  • Leppänen VM; Department of Biochemistry and Food Technology, University of Turku, Finland.
Protein Sci ; 8(6): 1218-31, 1999 Jun.
Article en En | MEDLINE | ID: mdl-10386872
The first crystal structure of an inorganic pyrophosphatase (S-PPase) from an archaebacterium, the thermophile Sulfolobus acidocaldarius, has been solved by molecular replacement and refined to an R-factor of 19.7% at 2.7 A. S-PPase is a D3 homohexameric protein with one Mg2+ per active site in a position similar to, but not identical with, the first activating metal in mesophilic pyrophosphatases (PPase). In mesophilic PPases, Asp65, Asp70, and Asp102 coordinate the Mg2+, while only Asp65 and Asp102 do in S-PPase, and the Mg2+ moves by 0.7 A. S-PPase may therefore be deactivated at low temperature by mispositioning a key metal ion. The monomer S-PPase structure is very similar to that of Thermus thermophilus (T-PPase) and Escherichia coli (E-PPase), root-mean-square deviations around 1 A/Calpha. But the hexamer structures of S- and T-PPase are more tightly packed and more similar to each other than they are to that of E-PPase, as shown by the increase in surface area buried upon oligomerization. In T-PPase, Arg116 creates an interlocking ionic network to both twofold and threefold related monomers; S-PPase has hydrophilic interactions to threefold related monomers absent in both E- and T-PPase. In addition, the thermostable PPases have about 7% more hydrogen bonds per monomer than E-PPase, and, especially in S-PPase, additional ionic interactions anchor the C-terminus to the rest of the protein. Thermostability in PPases is thus due to subtle improvements in both monomer and oligomer interactions.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Pirofosfatasas / Sulfolobus acidocaldarius / Metales Tipo de estudio: Prognostic_studies Idioma: En Revista: Protein Sci Asunto de la revista: BIOQUIMICA Año: 1999 Tipo del documento: Article País de afiliación: Finlandia Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Pirofosfatasas / Sulfolobus acidocaldarius / Metales Tipo de estudio: Prognostic_studies Idioma: En Revista: Protein Sci Asunto de la revista: BIOQUIMICA Año: 1999 Tipo del documento: Article País de afiliación: Finlandia Pais de publicación: Estados Unidos