The human metallochaperone HAH1 has been produced in Escherichia coli with four additional amino acids at the C-terminus and characterized in solution by NMR spectroscopy, both with and without copper(I). The solution structure of the apo-HAH1 monomer has a root-mean-square-deviation (RMSD) of 0.50 Å for the coordinates of the backbone atoms and 0.96 Å for all heavy atoms. These values compare, respectively, with 0.45 and 0.95 Å for copper(I)-HAH1. There are only minor structural rearrangements upon copper(I) binding. In particular, the variation of interatomic interactions around the metal-binding region is limited to a movement of Lys60 toward the metal site. The protein structures are similar to those obtained by X-ray crystallography in a variety of derivatives, with backbone RMSD values below 1 Å. In the holoprotein, copper(I) is confirmed to be two coordinated. If these data are compared with those of orthologue proteins, we learn that HAH1 has a lower tendency to change coordination number from two to three. Such a switch in coordination is a key step in copper transfer.
Solution structure of the apo and copper(I)-loaded human metallo-chaperone HAH1 / J.Anastassopoulou; L.Banci; I.Bertini; F.Cantini; E.Katsari; A.Rosato. - In: BIOCHEMISTRY. - ISSN 0006-2960. - STAMPA. - 43:(2004), pp. 13046-13053. [10.1021/bi0487591]
Solution structure of the apo and copper(I)-loaded human metallo-chaperone HAH1
BANCI, LUCIA;BERTINI, IVANO;CANTINI, FRANCESCA;ROSATO, ANTONIO
2004
Abstract
The human metallochaperone HAH1 has been produced in Escherichia coli with four additional amino acids at the C-terminus and characterized in solution by NMR spectroscopy, both with and without copper(I). The solution structure of the apo-HAH1 monomer has a root-mean-square-deviation (RMSD) of 0.50 Å for the coordinates of the backbone atoms and 0.96 Å for all heavy atoms. These values compare, respectively, with 0.45 and 0.95 Å for copper(I)-HAH1. There are only minor structural rearrangements upon copper(I) binding. In particular, the variation of interatomic interactions around the metal-binding region is limited to a movement of Lys60 toward the metal site. The protein structures are similar to those obtained by X-ray crystallography in a variety of derivatives, with backbone RMSD values below 1 Å. In the holoprotein, copper(I) is confirmed to be two coordinated. If these data are compared with those of orthologue proteins, we learn that HAH1 has a lower tendency to change coordination number from two to three. Such a switch in coordination is a key step in copper transfer.I documenti in FLORE sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.