[Analyzing secondary structure of beta2 microglobulin in Escherichia coli with circular dichroism spectrum].
OBJECTIVE
In order to study the structure and function of beta2 microglobulin (beta2 m).


METHODS
We sub-cloned the mature peptide of beta2 m into the p2X plasmid and transformed them to Escherichia coli TB1. The recombinant bacterium was induced to be expressed and the expressed fusion protein was detected by SDS-PAGE and western blot. After purifying and cleaving with Factor Xa, we separated the monomer protein beta2 m from MBP. Finally, we determined the secondary structure of the beta2 m protein by circular dichroism (CD) spectrum.


RESULTS
MBP-beta2 m was 52.1 kDa, and the monomer protein beta2 m was 10.6 kDa. The alpha-helix, beta-sheet, turn, and random coil of the fusion protein showed 0, 45, 8 and 45 amino acids, respectively, detected by CD estimation.


CONCLUSIONS
The beta2 m protein had correct secondary structure and could be used for further research of peptide binding in vitro.
Gao
Fengshan
F
Dalian University, Bioengineering College, Dalian 116622, China. gfsh0626@126.com
Xia
Chun
C
Zhang
Qiang
Q
Li
Yungang
Y
Li
Xinsheng
X
Xu
Chongbo
C
Bai
Jing
J
D000818:Animals; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D008970:Molecular Weight; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013552:Swine; D001613:beta 2-Microglobulin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D001613:beta 2-Microglobulin