Construction and characterization of a chimeric myoglobin

• Published in: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics
• Main Authors: Raffaella Musto, BRUNORI, Maurizio, CUTRUZZOLA', Francesca, TRAVAGLINI ALLOCATELLI, Carlo
• Other Authors: Raffaella, Musto, Brunori, Maurizio, Cutruzzola', Francesca
• Format: Article in Journal/Newspaper
• Language: English
• Published: ELSEVIER SCIENCE BV 2003
• Subjects: aplysia-limacina myoglobin; chimera; folding; folding intermediate; molten globule; myoglobin; reactivity; Sperm whale
• Online Access: http://hdl.handle.net/11573/254858; https://doi.org/10.1016/s1570-9639(02)00528-9; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000180957900004&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=0c7ff228ccbaaa74236f48834a34396a; http://www.scopus.com/inward/record.url?eid=2-s2.0-1242341185&partnerID=65&md5=b6f38c20e2854398fba201899e702538

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.