[Escherichia coli disulfide-forming related proteins: structures, functions and their application in gene engineering for expressing heterologous proteins in Escherichia coli].
The formation of disulfide bonds in secreted proteins of E. coli is a synergetic process depending on a series of Dsb proteins containing DsbA, DsbB, DsbC, DsbD, DsbE and DsbG. DsbA functions as an oxidant to form a disulfide bond between two -SH- in vivo and DsbB reactivates DsbA by reoxidizing it. Both DsbC and DsbG, two periplasmic proteins with isomerase activity, can correct mis-paired disulfide bonds introduced by DsbA although they recognize different substrates. DsbD, an inner membrane protein, plays a role in reducing DsbC and DsbG in vivo. It is regarded that DsbE has the similar function with DsbD. All DsbA, DsbC and DsbG have chaperone activity besides involving in the formation of disulfide bonds. Furthermore, their chaperone activity can promote the formation of protein disulfide bonds. There are a few reports dealing with soluble expression of heterologous proteins containing disulfide bonds assisted by DsbA and DsbC in E. coli. So far there has been no reports about the soluble expression of heterologous proteins promoted by DsbG. Our experiments first demonstrated that both DsbC and DsbG can improve the expression of single chain antibodies as soluble and functional forms in E. coli, and DsbG has additive effects with DsbC.
Zhang
Zhong
Z
Institute of Genetics, Chinese Academy of Sciences, Beijing 100101, China.
Huang
Hua-Liang
HL
D001426:Bacterial Proteins / Q000737:chemistry / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005818:Genetic Engineering / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry / Q000502:physiology*; D018832:Molecular Chaperones / Q000502:physiology; D010088:Oxidoreductases / Q000737:chemistry / Q000502:physiology*; D033903:Periplasmic Proteins; D019704:Protein Disulfide-Isomerases / Q000737:chemistry / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis*
D001426:Bacterial Proteins; C079503:DsbB protein, Bacteria; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D018832:Molecular Chaperones; D033903:Periplasmic Proteins; D011994:Recombinant Proteins; D010088:Oxidoreductases; C109831:DsbG protein, E coli; D019704:Protein Disulfide-Isomerases