In order to investigate the functional and structural role of modular structure in globins, we have engineered a chimeric myoglobin (ChimMb) in which the first and third exon come from the gene coding for the sperm whale Mb and the second exon from the gene coding for Aplysia limacina Mb. This ChimMb, fused to the Maltose Binding Protein (MBP) and expressed in Escherichia coli as an apoprotein, binds protoheme in a 1:1 stoichiometric ratio. Based on some functional and spectroscopic properties, we conclude that the central core of the ChimMb (which derives from A. limacina) is native-like. On the other hand, the ChimMb deprived (by proteolytic digestion) of the fused MBP displays a considerably reduced stability. These results suggest that the sperm whale A-G-H nucleus does not contribute significantly to the overall stability of the ChimMb. (C) 2002 Elsevier Science B.V. All rights reserved.
Construction and characterization of a chimeric myoglobin / Raffaella, Musto; Brunori, Maurizio; Cutruzzola', Francesca; TRAVAGLINI ALLOCATELLI, Carlo. - In: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS. - ISSN 1570-9639. - 1645:2(2003), pp. 139-145. [10.1016/s1570-9639(02)00528-9]
Construction and characterization of a chimeric myoglobin
BRUNORI, Maurizio;CUTRUZZOLA', Francesca;TRAVAGLINI ALLOCATELLI, Carlo
2003
Abstract
In order to investigate the functional and structural role of modular structure in globins, we have engineered a chimeric myoglobin (ChimMb) in which the first and third exon come from the gene coding for the sperm whale Mb and the second exon from the gene coding for Aplysia limacina Mb. This ChimMb, fused to the Maltose Binding Protein (MBP) and expressed in Escherichia coli as an apoprotein, binds protoheme in a 1:1 stoichiometric ratio. Based on some functional and spectroscopic properties, we conclude that the central core of the ChimMb (which derives from A. limacina) is native-like. On the other hand, the ChimMb deprived (by proteolytic digestion) of the fused MBP displays a considerably reduced stability. These results suggest that the sperm whale A-G-H nucleus does not contribute significantly to the overall stability of the ChimMb. (C) 2002 Elsevier Science B.V. All rights reserved.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
