Cloning, expression and purification of Atlantic salmon (Salmo salar, L.) neuroglobin

Neuroglobin (Ngb) exists only in small amounts in salmon brain. In order to study the protein in more detail salmon neuroglobin (sNgb) was cloned, hetereologously expressed in E.coli and purified. The protein had red color and showed the characteristic peaks at 411 nm (metNgb), 415 nm (carboxyNgb) a...

Full description

Bibliographic Details
Published in:Protein Expression and Purification
Main Authors: Bjørlykke, Gry Aletta, Kvamme, Bjørn Olav, Slinde, Erik, Raae, Arnt Johan
Format: Article in Journal/Newspaper
Language:English
Published: Elsevier Inc. 2012
Subjects:
Neuroglobin
Atlantic salmon
western analysis
heme spectrum
homology model
psycrophilic
Salmo salar
Online Access:https://hdl.handle.net/1956/6219
https://doi.org/10.1016/j.pep.2012.09.010
Description
Summary:Neuroglobin (Ngb) exists only in small amounts in salmon brain. In order to study the protein in more detail salmon neuroglobin (sNgb) was cloned, hetereologously expressed in E.coli and purified. The protein had red color and showed the characteristic peaks at 411 nm (metNgb), 415 nm (carboxyNgb) and 424 nm (deoxyNgb). Western analysis showed that sNgb reacted weakly against a rabbit anti human neuroglobin (hNgb) and strongly to a sNgb specific antibody. Our 3Dhomology model of the sNgb indicated modifications adjacent to and in the O2/CO binding site. This may correlate to differences in substrate affinities for the sNgb compared to the hNgb. Also sNgb contained shorter helixes and longer interhelical loops typical for psycrophilic proteins. submittedVersion

Similar Items